| identification of ribonucleotide reductase protein r1 as an activator of microtubule nucleation in xenopus egg mitotic extracts. | microtubule nucleation on the centrosome and the fungal equivalent, the spindle pole body (spb), is activated at the onset of mitosis. we previously reported that mitotic extracts prepared from xenopus unfertilized eggs convert the interphase spb of fission yeast into a competent state for microtubule nucleation. in this study, we have purified an 85-kda spb activator from the extracts and identified it as the ribonucleotide reductase large subunit r1. we further confirmed that recombinant mouse ... | 2000 | 11102516 |
| construction and characterization of bispecific costimulatory molecules containing a minimized cd86 (b7-2) domain and single-chain antibody fragments for tumor targeting. | efficient t-cell activation requires two signals. the first signal, which confers specificity, is provided by interaction of the t-cell receptor with peptides presented by mhc molecules. one of the second costimulatory signals is induced by binding of b7 proteins on the surface of antigen-presenting cells to cd28 on the t-cell surface. expression of b7 molecules on tumor cells can result in the activation of tumor specific t lymphocytes and induce protective antitumor immunity. however, at prese ... | 2000 | 11106249 |
| the mouse spo11 gene is required for meiotic chromosome synapsis. | the spo11 protein initiates meiotic recombination by generating dna double-strand breaks (dsbs) and is required for meiotic synapsis in s. cerevisiae. surprisingly, spo11 homologs are dispensable for synapsis in c. elegans and drosophila yet required for meiotic recombination. disruption of mouse spo11 results in infertility. spermatocytes arrest prior to pachytene with little or no synapsis and undergo apoptosis. we did not detect rad51/dmc1 foci in meiotic chromosome spreads, indicating dsbs a ... | 2000 | 11106738 |
| cloning and characterization of human vps35 and mouse vps35 and mapping of vps35 to human chromosome 16q13-q21. | maintenance of different organelles in eukaryotic cells depends on sorting proteins, which ensure the proper delivery of organelle-specific proteins. the studies on yeast (saccharomyces cerevisiae) vps35, a hydrophilic membrane protein having a direct role in the retrieval of cargo proteins, suggest a mechanism underlying a possible lysosomal protein-sorting pathway in mammalian cells. here, we report the isolation of human and mouse vps35 cdnas, which are 3208 and 3186 bp in length, respectivel ... | 2000 | 11112353 |
| distinct regions of the chicken p46 polypeptide are required for its in vitro interaction with histones h2b and h4 and histone acetyltransferase-1. | we cloned cdna encoding the chicken p46 polypeptide, chp46, homologous to the p48 subunit of chicken chromatin assembly factor-1, chcaf-1p48. it comprises 424 amino acids including a putative initiation met, is a member of the wd protein family, with seven wd repeat motifs, and exhibits 90.3% identity to chcaf-1p48 and 94.3% identity to the human and mouse p46 polypeptides (hup46 and mop46). the in vitro immunoprecipitation experiment established that chp46 interacts with histones h2b and h4 and ... | 2000 | 11112423 |
| msx3 protein recruits histone deacetylase to down-regulate the msx1 promoter. | msx1 promoter is known to be repressed by msx1 protein [shetty, takahashi, matsui, iyengar and raghow (1999) biochem. j. 339, 751-758]. we show that in the transiently transfected c(2)c(12) myoblasts, co-expression of msx3 also causes potent repression of msx1 promoter that can be relieved by exogenous expression of camp-response-element-binding protein-binding protein (cbp) and p300 in a dose-dependent manner. co-immunoprecipitation and western blot analyses revealed that msx3 interacts with cb ... | 2001 | 11115394 |
| involvement of the tom complex in external nadh transport into yeast mitochondria depleted of mitochondrial porin1. | the protein(s) responsible for metabolite transport through the outer membrane of the yeast saccharomyces cerevisiae mitochondria depleted of mitochondrial porin (also known as voltage-dependent anion selective channel), termed here porin1, is (are) still unidentified. it is postulated that the transport may be supported by the protein import machinery of the outer membrane, the tom complex (translocase of the outer membrane). we demonstrate here that in the absence of functional porin1, the blo ... | 2000 | 11118520 |
| expression of the protein product of the pcph proto-oncogene in human tumor cell lines. | expression of the protein product of the pcph proto-oncogene in human tumor cell lines. exposure of syrian hamster embryo fibroblasts to chemical carcinogens resulted in the oncogenic activation of the pcph proto-oncogene by induction of a single base-pair deletion that generated a truncated pcph oncoprotein (mutated pcph). recently, we isolated and characterized the cdna for the human pcph proto-oncogene and determined that in humans pcph is a single-copy gene located in chromosome 14 (14q24.3) ... | 2001 | 11121232 |
| interaction of the c-terminal tail region of the metabotropic glutamate receptor 7 with the protein kinase c substrate pick1. | group iii metabotropic glutamate receptors (mglurs) are highly enriched in the presynaptic terminals of glutamatergic synapses where they mediate feedback inhibition of neurotransmitter release. here, we used the yeast two-hybrid system to identify a direct interaction of the c-terminal tail region of mglur7 with the rat homologue of the protein kinase c substrate pick1. this interaction is specifically mediated by the very c-terminal amino acids of the receptor and can be reconstituted in human ... | 2000 | 11122333 |
| candidate mutator genes in mismatch repair-deficient thymic lymphomas: no evidence of mutations in the dna polymerase delta gene. | dna mismatch repair (mmr) proteins recognize nucleotides that are incorrectly paired. deficiencies in mmr lead to increased genomic instability reflected in an increased mutation frequency and predisposition to tumorigenesis. mice lacking the mmr gene, msh2, develop thymic lymphomas that exhibit much higher mutational frequencies than other msh2(-/-) tumours and msh2(-/-) normal thymic tissue, suggesting that an additional mutator may have been acquired in a tissue-specific manner. clustered mut ... | 2000 | 11133819 |
| heterozygous hesx1 mutations associated with isolated congenital pituitary hypoplasia and septo-optic dysplasia. | we have previously shown that familial septo-optic dysplasia (sod), a syndromic form of congenital hypopituitarism involving optic nerve hypoplasia and agenesis of midline brain structures, is associated with homozygosity for an inactivating mutation in the homeobox gene hesx1/hesx1 in man and mouse. however, as most sod/congenital hypopituitarism occurs sporadically, the possible contribution of hesx1 mutations to the aetiology of these cases is presently unclear. interestingly, a small proport ... | 2001 | 11136712 |
| a 5-bp deletion in elovl4 is associated with two related forms of autosomal dominant macular dystrophy. | stargardt-like macular dystrophy (stgd3, mim 600110) and autosomal dominant macular dystrophy (admd) are inherited forms of macular degeneration characterized by decreased visual acuity, macular atrophy and extensive fundus flecks. genetic mapping data suggest that mutations in a single gene may be responsible for both conditions, already known to bear clinical resemblance. here we limit the minimum genetic region for stgd3 and admd to a 0.6-cm interval by recombination breakpoint mapping and id ... | 2001 | 11138005 |
| characterization of mouse ire1 alpha: cloning, mrna localization in the brain and functional analysis in a neural cell line. | in yeast, an endoplasmic reticulum (er)-associated protein, ire1p, is believed to initiate the unfolded protein response (upr), that is responsible for protein folding in the er under stressed conditions. two mammalian homologs of ire1p have been identified, ire1 alpha and ire1 beta. we have previously reported that familial alzheimer's disease linked presenilin-1 variants downregulate the signaling pathway of the upr by affecting the phosphorylation of ire1 alpha. in the present study, we clone ... | 2000 | 11146108 |
| direct interactions of the five known fanconi anaemia proteins suggest a common functional pathway. | fanconi anaemia (fa) is an autosomal recessive inherited disorder associated with a progressive aplastic anaemia, diverse congenital abnormalities and cancer. the condition is genetically heterogeneous, with at least seven complementation groups (a-g) described. cells from individuals who are homozygous for mutations in fa genes are characterized by chromosomal instability and hypersensitivity to dna interstrand crosslinking agents. these features suggest a possible role for the encoded proteins ... | 2001 | 11157805 |
| two distinct nuclear receptor-interaction domains and creb-binding protein-dependent transactivation function of activating signal cointegrator-2. | asc-2 is a recently isolated transcriptional cointegrator molecule, which is amplified in human cancers and stimulates transactivation by nuclear receptors, ap-1, nuclear factor kappab (nfkappab), serum response factor (srf), and numerous other transcription factors. asc-2 contained two nuclear receptor-interaction domains, both of which are dependent on the integrity of their core lxxll sequences. surprisingly, the c-terminal lxxll motif specifically interacted with oxysterol receptor lxrss, wh ... | 2001 | 11158331 |
| n-linked oligosaccharide processing enzyme glucosidase ii produces 1,5-anhydrofructose as a side product. | alpha-1,4-glucan lyase cleaves alpha-1,4-linkages of nonreducing termini of alpha-1,4-glucans to produce 1,5-anhydrofructose (1,5-anfru). the enzymes isolated from fungi and algae show high homology with glycoside hydrolase family 31. purification of alpha-1,4-glucan lyase from rat liver using deae cellulose chromatography resulted in separation of two enzymatic active fractions, one was bound to the column and the other was in the flow-through. partial amino acid sequence determined from the ly ... | 2000 | 11159920 |
| determinants of human apolipoprotein [a] secretion from mouse hepatocyte cultures. | efforts to develop an in vitro model system to analyze apolipoprotein [a] (apo[a]) gene transcription, mrna translation, and protein secretion have been complicated by the limited tissue and species distribution of apo[a] and the presence of regulatory dna sequences remote from the apo[a] transcription start site. in the current study we examined primary hepatocytes cultured from apo[a] transgenic mice as a model system for analyzing apo[a] biogenesis. hepatocytes from mice transgenic for a yeas ... | 2001 | 11160366 |
| cloning and characterization of a potential transcriptional activator of human gamma-globin genes. | hybrids produced by fusing human fetal erythroblasts (hfe) with mouse erythroleukemia (mel) cells initially produce predominantly or exclusively human gamma-globin and switch to human beta globin expression as time in culture advances. one explanation for the initially predominant expression of gamma-globin gene in these hybrids is the presence of trans-acting factors that activate gamma-globin gene transcription. we used differential display of hybrids before and after the gamma to beta switch ... | 2001 | 11162141 |
| the double-stranded rna-activated protein kinase pkr is dispensable for regulation of translation initiation in response to either calcium mobilization from the endoplasmic reticulum or essential amino acid starvation. | the alpha-subunit of eukaryotic initiation factor eif2 is a preferred substrate for the double-stranded rna-activated protein kinase, pkr. phosphorylation of eif2alpha converts the factor from a substrate into a competitive inhibitor of the guanine nucleotide exchange factor, eif2b, leading to a decline in mrna translation. early studies provided evidence implicating pkr as the kinase that phosphorylates eif2alpha under conditions of cell stress such as the accumulation of misfolded proteins in ... | 2001 | 11162513 |
| two isoforms of trimming glucosidase ii exist in mammalian tissues and cell lines but not in yeast and insect cells. | we previously cloned glucosidase ii and provided in vivo evidence for its involvement in protein folding quality control. dna-sequencing of different clones demonstrated the existence of two isoforms of glucosidase ii which differed by 66 nucleotides due to alternative splicing. the existence of two enzyme isoforms in various organs of pig and rat as well as human, bovine, rat, and mouse cell lines could be demonstrated by rt-pcr and western blotting. furthermore, the two isoforms of glucosidase ... | 2001 | 11162524 |
| msiglec-e, a novel mouse cd33-related siglec (sialic acid-binding immunoglobulin-like lectin) that recruits src homology 2 (sh2)-domain-containing protein tyrosine phosphatases shp-1 and shp-2. | the sialic acid-binding immunoglobulin-like lectins (siglecs) represent a recently defined distinct subset of the immunoglobulin superfamily. by using the src homology 2 (sh2)-domain-containing protein tyrosine phosphatase shp-1 as bait in a yeast two-hybrid screen, we have identified a new member of the mouse siglec family, msiglec-e. the msiglec-e cdna encodes a protein of 467 amino acids that contains three extracellular immunoglobulin-like domains, a transmembrane region and a cytoplasmic ta ... | 2001 | 11171044 |
| characterization of sphingolipid long-chain base kinase in arabidopsis thaliana. | sphingolipid long-chain base (lcb) kinase catalyses the phosphorylation of sphingolipid lcb to form lcb 1-phosphate. based on sequence identity to a murine sphingosine kinase (murine sphk1a), we isolated and characterized a lcb kinase-like cdna in arabidopsis thaliana. the deduced amino acid sequence of the homologous cdna shows several regions that are highly conserved in lcb kinases from mouse, yeast, human and caenorhabditis elegans. these regions are not similar to those of other known kinas ... | 2000 | 11171192 |
| a fyve-finger-containing protein, rabip4, is a rab4 effector involved in early endosomal traffic. | the small gtpase rab4 is implicated in endocytosis in all cell types, but also plays a specific role in some regulated processes. to better understand the role of rab4 in regulation of vesicular trafficking, we searched for an effector(s) that specifically recognizes its gtp-bound form. we cloned a ubiquitous 69-kda protein, rabip4, that behaves as a rab4 effector in the yeast two-hybrid system and in the mammalian cell. rabip4 contains two coiled-coil domains and a fyve-finger domain. when expr ... | 2001 | 11172003 |
| characterization of promoter region and genomic structure of the murine and human genes encoding src like adapter protein. | src-like adapter protein (slap) was identified as a signaling molecule in a yeast two-hybrid system using the cytoplasmic domain of epha2, a receptor protein tyrosine kinase (pandey et al., 1995. characterization of a novel src-like adapter protein that associates with the eck receptor tyrosine kinase. j. biol. chem. 270, 19201-19204). it is very similar to members of the src family of cytoplasmic tyrosine kinases in that it contains very homologous sh3 and sh2 domains (abram and courtneidge, 20 ... | 2001 | 11179692 |
| extra-chromosomal telomeric dna in cells from atm(-/-) mice and patients with ataxia-telangiectasia. | ataxia-telangiectasia (at) is an autosomally recessive human genetic disease with pleiotropic defects such as neurological degeneration, immunodeficiency, chromosomal instability, cancer susceptibility and premature aging. cells derived from at patients and ataxia-telangiectasia mutated (atm)-deficient mice show slow growth in culture and premature senescence. atm, which belongs to the pi3 kinase family along with dna-pk, plays a major role in signaling the p53 response to dna strand breaks. tel ... | 2001 | 11181576 |
| calcium/calmodulin-dependent protein kinase ii inhibitor protein: localization of isoforms in rat brain. | a second isoform of ca2+/calmodulin-dependent-kinase ii inhibitor protein (cam-kiin) has been identified using the yeast two-hybrid screen. the 1.8kb message encodes a 78 residue cam-kiinalpha that is 65% identical in its putative open-reading frame and 95% identical in its inhibitory domain to the previously characterized cam-kiinbeta. cam-kiinalpha exhibits inhibitory properties towards recombinant mouse cam-kinase iialpha indistinguishable from cam-kiinbeta. the 27 amino acid inhibitory pepti ... | 2001 | 11182241 |
| expression and catalytic activity of mouse leukotriene b4 omega-hydroxylase, cyp4f14. | we have isolated a cdna for a mouse leukotriene b4 omega-hydroxylase, cyp4f14. the cdna encoded a protein with 524 amino acids, whose sequence similarity is 95% that of rat cyp4f1. the microsomes from yeast cells transfected with cyp4f14 expression vector showed 0.1 nmol p450/mg protein and catalyzed omega-hydroxylations of leukotriene b4, 6-trans-leukotriene b4, lipoxin a4, prostaglandin a1, and several hydroxyeicosatetraeonic acids (hetes), with 8-hete being the most active substrate. in contr ... | 2000 | 11185557 |
| evidence for the binding of a human sperm component with diaphanous protein. | the ywk-ii component of human sperm membrane is related to the betaa4-amyloid precursor protein (app) of alzheimer's disease. a yeast 2-hybrid system was used to screen a mouse testis cdna expression library for potential ligands capable of interacting with the extracellular domain of the ywk-ii component. one of the bound proteins was identified as hdia1, which has 96% identity with p140mdia. these proteins are members of the formin homology family and participate in cytokinesis and organizatio ... | 2001 | 11204614 |
| smcy transgene does not rescue spermatogenesis in sex-reversed mice. | in mouse, the sxr(b) deletion interval (delta sxr(b)) maps to the small short arm of the y chromosome and is known to contain gene(s) required for normal spermatogenesis; in particular, spy, which is essential for the postnatal mitotic proliferation of spermatogonia. this deletion interval is approximately 1-2 mb and contains eight known genes. in this paper we report the construction of yac transgenic mice containing different regions of the delta sxr(b) interval including zfy1, ube1y, smcy, an ... | 2001 | 11210179 |
| identification of mouse jun dimerization protein 2 as a novel repressor of atf-2. | a mouse cdna that encodes a dna-binding protein was identified by yeast two-hybrid screening, using activating transcription factor-2 (atf-2) as the bait. the protein contained a bzip (basic amino acid-leucine zipper region) domain and its amino acid sequence was almost identical to that of rat jun dimerization protein 2 (jdp2). mouse jdp2 interacted with atf-2 both in vitro and in vivo via its bzip domain. it was encoded by a single gene and various transcripts were expressed in all tested tiss ... | 2001 | 11231009 |
| phosphorylation and rapid relocalization of 53bp1 to nuclear foci upon dna damage. | 53bp1 is a human brct protein that was originally identified as a p53-interacting protein by the saccharomyces cerevisiae two-hybrid screen. although the carboxyl-terminal brct domain shows similarity to crb2, a dna damage checkpoint protein in fission yeast, there is no evidence so far that implicates 53bp1 in the checkpoint. we have identified a xenopus homologue of 53bp1 (xl53bp1). xl53bp1 is associated with chromatin and, in some cells, localized to a few large foci under normal conditions. ... | 2001 | 11238909 |
| the small gtpase rab4a interacts with the central region of cytoplasmic dynein light intermediate chain-1. | rab4 belongs to the rab family of small gtpases involved in the regulation of intracellular transport, and has been localized to early endosomes. we have employed the yeast two-hybrid system to identify proteins that specifically interact with rab4aq67l, a gtpase-deficient mutant form of rab4a. screening a mouse embryo cdna library identified a clone (m449) that interacted with rab4a in a nucleotide-dependent fashion. data base searches identified this clone as the mouse cytoplasmic dynein light ... | 2001 | 11243854 |
| characterization of mouse brain-specific angiogenesis inhibitor 1 (bai1) and phytanoyl-coa alpha-hydroxylase-associated protein 1, a novel bai1-binding protein. | previously, pahx-ap1 (pahx-associated protein 1) was isolated as a novel protein to interact with refsum disease gene product (phytanoyl-coa alpha-hydroxylase, pahx) and specifically expressed in mouse brain. pahx-ap1 is also suggested to be involved in the development of the central neurologic deficits of refsum disease. to clarify its function, we have searched for proteins that associate with pahx-ap1 via yeast two-hybrid system. we found that pahx-ap1 interacts with the cytoplasmic region of ... | 2001 | 11245925 |
| a double-deletion mutation in the pitx3 gene causes arrested lens development in aphakia mice. | the recessive aphakia (ak) mouse mutant is characterized by bilateral microphthalmia due to a failure of lens morphogenesis. we fine-mapped the ak locus to the interval between d19umi1 and d19mit9, developed new polymorphic markers, and mapped candidate genes by construction of a bac contig. the pitx3 gene, known to be expressed in lens primordia, shows zero recombination with the ak mutation on our intersubspecific intercross panel representing 1170 meioses. a recent report described a deletion ... | 2001 | 11247667 |
| the suppressor of fused gene, involved in hedgehog signal transduction in drosophila, is conserved in mammals. | the suppressor of fused [su(fu)] gene of drosophila melanogaster encodes a protein containing a pest sequence [sequence enriched in proline (p), glutamic acid (e), serine (s) and threonine (t)] which acts as an antagonist to the serine-threonine kinase fused in hedgehog (hh) signal transduction during embryogenesis. the su(fu) gene isolated from a distantly related drosophila species, d. virilis, shows significantly high homology throughout its protein sequence with its d. melanogaster counterpa ... | 1999 | 11252182 |
| elevated expression of sag/roc2/rbx2/hrt2 in human colon carcinomas: sag does not induce neoplastic transformation, but antisense sag transfection inhibits tumor cell growth. | sensitive-to-apoptosis gene (sag)/regulator of cullins (roc)2/rbx2/hrt2 is a newly identified component of scf e3 ubiquitin ligase that controls cell-cycle progression by promoting ubiquitination and degradation of cell-cycle inhibitors. we recently found that sag protects cells from apoptosis induced by redox agents, promotes s-phase entry and cell growth under serum starvation, and is required for yeast growth. in the present study, we report that the sag protein level was elevated in six of 1 ... | 2001 | 11255265 |
| sez4 gene encoding an elongation subunit of dna polymerase zeta is required for normal embryogenesis. | sez4 identified as a seizure-activated gene shows a similarity to the yeast rev3 that encodes a catalytic subunit of the nonessential dna polymerase zeta which is involved in error-prone translesion synthesis. although yeast rev3 homologues in mouse and human have recently been identified and characterized, their precise roles remain elusive. | 2001 | 11260255 |
| a metabolic enzyme for s-nitrosothiol conserved from bacteria to humans. | considerable evidence indicates that no biology involves a family of no-related molecules and that s-nitrosothiols (snos) are central to signal transduction and host defence. it is unknown, however, how cells switch off the signals or protect themselves from the snos produced for defence purposes. here we have purified a single activity from escherichia coli, saccharomyces cerevisiae and mouse macrophages that metabolizes s-nitrosoglutathione (gsno), and show that it is the glutathione-dependent ... | 2001 | 11260719 |
| development of two bacterial artificial chromosome shuttle vectors for a recombination-based cloning and regulated expression of large genes in mammalian cells. | most conditional expression vectors designed for mammalian cells have been valuable systems for studying genes of interest by regulating their expressions. the available vectors, however, are reliable for the short-length cdna clones and not optimal for relatively long fragments of genomic dna or long cdnas. here, we report the construction of two bacterial artificial chromosome (bac) vectors, capable of harboring large inserts and shuttling among escherichia coli, yeast, and mammalian cells. th ... | 2001 | 11262167 |
| of yeast, mice, and men. rab proteins and organelle transport. | | 2001 | 11266477 |
| cloning and functional characterization of chicken p160 coactivator family members. | the factors src-1, tif2 and actr were identified as interacting with nuclear receptors in a highly ligand-dependent manner. because the molecular mass of each of these factors is approximately 160 kda, they are collectively termed p160 coactivators. so far, p160 coactivators have been cloned from human, mouse and xenopus. we report here the cloning of the chicken homologues of p160 coactivator members. as in human and mouse, chicken has three p160 coactivators. each gene encodes an approximately ... | 2001 | 11267654 |
| isolation and characterization of cdnas for the protein kinase hipk2. | hipk2 (homeodomain-interacting protein kinase 2) is a cd95 binding partner in yeast. its primary amino acid sequence is highly conserved between human and mouse. the highest hipk2 mrna expression is found in neuronal tissue. the hipk2 gene is located on human chromosome 7q33-35 and the protein is mainly localized in the nucleus. hipk2 has been described to play a role as a co-repressor for homeodomain transcription factors. | 2001 | 11267674 |
| augmenting transgene expression from carcinoembryonic antigen (cea) promoter via a gal4 gene regulatory system. | though extensively studied, the use of tissue- or cell-type-specific promoters to target transgene expression is hampered by their weak activity. we hypothesized that this problem could be addressed by using a gal4 gene regulatory system, wherein a weak, tissue-specific promoter would drive expression of the gal4/vp16 fusion protein (gv16), which in turn would transactivate a minimal synthetic promoter, gal4/tata (gt), upstream of a transgene. to test this hypothesis, we constructed adenoviral v ... | 2001 | 11273768 |
| a splicing factor, prp8: preferential localization in the testis and ovary in adult mice. | prp8 is a splicing factor of 220 kda originally identified in yeast and is a component of the u5 small nuclear ribonucleoprotein particle. mouse prp8 cdna was cloned and shown to share 62.6 and 68.2% sequence identity with the yeast homologue at the amino acid and nucleotide level, respectively, while it differs by only 3 amino acid residues from the human homologue. during mouse embryogenesis, prp8 is expressed intensely at day 9.5 of gestation, and its expression decreases progressively during ... | 2001 | 11275560 |
| determination of protein-protein interactions of icin by the yeast two-hybrid system. | icln is a ubiquitously expressed eukaryotic protein. expression of the protein in xenopus laevis oocytes, the knocking-down of the protein in fibroblasts, or the reconstitution of the protein in lipid bilayer led to the assumption that this protein is an ionic channel or a significant part thereof. however, other possible roles for icln in potential regulatory mechanisms have been postulated, as diverse as regulator of cell morphology by interacting with the skb1 protein and/or interaction with ... | 2001 | 11275683 |
| the human tumor suppressor arf interacts with spinophilin/neurabin ii, a type 1 protein-phosphatase-binding protein. | the ink4a gene, one of the most often disrupted loci in human cancer, encodes two unrelated proteins, p16(ink4a) and p14(arf) (arf) both capable of inducing cell cycle arrest. although it has been clearly demonstrated that arf inhibits cell cycle via p53 stabilization, very little is known about the involvement of arf in other cell cycle regulatory pathways, as well as on the mechanisms responsible for activating arf following oncoproliferative stimuli. in search of factors that might associate ... | 2001 | 11278317 |
| trax (translin-associated factor x), a primarily cytoplasmic protein, inhibits the binding of tb-rbp (translin) to rna. | trax (translin-associated factor x) has been shown to interact with tb-rbp/translin by its coimmunoprecipitation and in yeast two-hybrid assays. here we demonstrate that trax is widely expressed, does not bind to dna or rna, but forms heterodimers with tb-rbp under reducing conditions. the heterodimer of tb-rbp and trax inhibits tb-rbp binding to rna, but enhances tb-rbp binding to specific single stranded dna sequences. the in vitro interactions between tb-rbp and trax are confirmed by similar ... | 2001 | 11278549 |
| mechanisms governing subcellular localization and function of human rgs2. | rgs proteins negatively regulate heterotrimeric g proteins at the plasma membrane. rgs2-gfp localizes to the nucleus, plasma membrane, and cytoplasm of hek293 cells. expression of activated g(q) increased rgs2 association with the plasma membrane and decreased accumulation in the nucleus, suggesting that signal-induced redistribution may regulate rgs2 function. thus, we identified and characterized a conserved n-terminal domain in rgs2 that is necessary and sufficient for plasma membrane localiz ... | 2001 | 11278586 |
| regulation of stra13 by the von hippel-lindau tumor suppressor protein, hypoxia, and the ubc9/ubiquitin proteasome degradation pathway. | in this study, we focus on different modes of regulation of stra13, a human ortholog of the mouse basic helix-loop-helix transcriptional factor, previously identified by us as a new von hippel-lindau tumor suppressor gene (vhl) target. the gene was overexpressed in vhl-deficient cell lines and tumors, specifically clear cell renal carcinomas and hemangioblastomas. introduction of wild type vhl transgene into clear cell renal carcinoma restored low level expression of stra13. overexpression was a ... | 2001 | 11278694 |
| dna binding by the ets-domain transcription factor pea3 is regulated by intramolecular and intermolecular protein.protein interactions. | the control of dna binding by eukaryotic transcription factors represents an important regulatory mechanism. many transcription factors are controlled by cis-acting autoinhibitory modules that are thought to act by blocking promiscuous dna binding in the absence of appropriate regulatory cues. here, we have investigated the determinants and regulation of the autoinhibitory mechanism employed by the ets-domain transcription factor, pea3. dna binding is inhibited by a module composed of a combinat ... | 2001 | 11278941 |
| the binding of ku antigen to homeodomain proteins promotes their phosphorylation by dna-dependent protein kinase. | the ku antigen (70- and 80-kda subunits) is a regulatory subunit of dna-dependent protein kinase (dna-pk) that promotes the recruitment of the catalytic subunit of dna-pk (dna-pkcs) to dna ends and to specific dna sequences from which the kinase is activated. ku and dna-pkcs plays essential roles in double-stranded dna break repair and v(d)j recombination and have been implicated in the regulation of specific gene transcription. in a yeast two-hybrid screen of a jurkat t cell cdna library, we ha ... | 2001 | 11279128 |
| isolation of 2000-kda complexes of n-methyl-d-aspartate receptor and postsynaptic density 95 from mouse brain. | neurotransmitter receptors in vivo are linked to intracellular adaptor proteins and signalling molecules driving downstream pathways. methods for physical isolation are essential to answer fundamental questions about the size, structure and composition of in vivo complexes and complement the widely used yeast 2-hybrid method. the n-methyl-d-aspartate receptor (nmdar) binds postsynaptic density 95 (psd-95) protein; both are required for synaptic plasticity and learning and participate in other im ... | 2001 | 11279284 |
| pathogenesis of prion diseases: a progress report. | almost 20 years have passed since stanley prusiner proposed that the agent causing transmissible spongiform encephalopathies consists exclusively of a protein and termed it prion. a mixed balance can be drawn from the enormous research efforts that have gone into prion research during this time. on the negative side, the protein-only hypothesis has not been conclusively proven yet. on the positive side, our understanding of spongiform encephalopathies has experienced tremendous advances, mostly ... | 2000 | 11279534 |
| regulation of eosinophil and neutrophil apoptosis--similarities and differences. | apoptosis is the most common form of physiologic cell death and a necessary process to maintain cell numbers in multicellular organisms. in many chronic inflammatory diseases, reduced cell death of different types of granulocytes is one important mechanism for cell accumulation. granulocytes are constantly produced in large amounts in the bone marrow and the same numbers die, under normal circumstances, within a defined time period. changing the rate of apoptosis rapidly changes cell numbers in ... | 2001 | 11292018 |
| its8, a fission yeast homolog of mcd4 and pig-n, is involved in gpi anchor synthesis and shares an essential function with calcineurin in cytokinesis. | in fission yeast, calcineurin is required for cytokinesis and ion homeostasis; however, most of its physiological roles remain obscure. to identify genes that share an essential function with calcineurin, we screened for mutations that confer sensitivity to the calcineurin inhibitor fk506 and high temperature and isolated the mutant its8-1. its8(+) encodes a homolog of the budding yeast mcd4 and human pig-n that are involved in glycosylphosphatidylinositol (gpi) anchor synthesis. consistently, r ... | 2001 | 11297516 |
| the cryptococcus neoformans ste11alpha gene is similar to other fungal mitogen-activated protein kinase kinase kinase (mapkkk) genes but is mating type specific. | partial sequence analysis of the cryptococcus neoformans matalpha mating type locus revealed the presence of a gene with substantial sequence similarity to other fungal mitogen-activated protein (map) kinase kinase kinase (mapkkk) genes. the c. neoformans gene, designated ste11alpha, showed the highest degree of similarity to the neurospora crassa nrc-1, schizosaccharomyces pombe byr2 and saccharomyces cerevisiae ste11 genes. a polymerase chain reaction-mediated sib-selection technique was succe ... | 2001 | 11298287 |
| tumor-associated antigens identified by mrna expression profiling induce protective anti-tumor immunity. | defined tumor-associated antigens (taa) are attractive targets for anti-tumor immunotherapy. here, we describe a novel genome-wide approach to identify multiple taa from any given tumor. a panel of transplantable thymomas was established from an inbred p53-/- mouse strain. the resulting tumors were examined for gene expression by mrna microarray scanning. this analysis revealed heterogeneity of the tumors in agreement with the assumption that they represent different tumorigenic events. several ... | 2001 | 11298350 |
| cloning, organisation, chromosomal localization and expression analysis of the mouse prkag1 gene. | the mammalian 5'-amp-activated protein kinase (ampk) is a heterotrimeric protein consisting of alpha-, beta- and gamma-subunits. the alpha-subunit is the catalytic subunit. the non-catalytic subunits ampk-beta and ampk-gamma form, together with the catalytic ampk-alpha, the active kinase complex in mammals and its homologue in yeast. the gene for ampk-gamma-1 has been designated recently as prkag1. we have isolated mouse prkag1 cdna from testis (1623 nt) coding for 330 aa and we have shown its u ... | 2001 | 11306812 |
| a novel snare n-terminal domain revealed by the crystal structure of sec22b. | intra-cellular membrane fusion is facilitated by the association of snares from opposite membranes into stable alpha-helical bundles. many snares, in addition to their alpha-helical regions, contain n-terminal domains that likely have essential regulatory functions. to better understand this regulation, we have determined the 2.4-a crystal structure of the 130-amino acid n-terminal domain of mouse sec22b (msec22b), a snare involved in endoplasmic reticulum/golgi membrane trafficking. the domain ... | 2001 | 11309394 |
| osp/claudin-11 forms a complex with a novel member of the tetraspanin super family and beta1 integrin and regulates proliferation and migration of oligodendrocytes. | oligodendrocyte-specific protein (osp)/claudin-11 is a major component of central nervous system myelin and forms tight junctions (tjs) within myelin sheaths. tjs are essential for forming a paracellular barrier and have been implicated in the regulation of growth and differentiation via signal transduction pathways. we have identified an osp/claudin-11-associated protein (oap)1, using a yeast two-hybrid screen. oap-1 is a novel member of the tetraspanin superfamily, and it is widely expressed i ... | 2001 | 11309411 |
| a novel actin-related protein gene of colletotrichum gloeosporioides f. sp. malvae shows altered expression corresponding with spore production. | a novel actin-related protein (arp) was found in the plant pathogenic fungus, colletotrichum gloeosporioides f. sp. malvae (cgm), which causes anthracnose disease of round-leaved mallow (malva pusilla). sequence comparisons showed that this gene, arpa, belongs to the highly divergent 'other arps' category in the current arp classification system. arpa is most similar to the arp11 gene of mus musculus but has a unique structure with deletions at the c-terminus similar to that of the arp10 gene of ... | 2001 | 11313136 |
| runx2: a novel oncogenic effector revealed by in vivo complementation and retroviral tagging. | the runx2 (cbfa1, pebp2alphaa, aml3) gene was previously identified as a frequent target for transcriptional activation by proviral insertion in t-cell lymphomas of cd2-myc transgenic mice. we have recently shown that over-expression of the full-length, most highly expressed runx2 isoform in the thymus perturbs t-cell development, leads to development of spontaneous lymphomas at low frequency and is strongly synergistic with myc. to gain further insight into the relationship of runx2 to other ly ... | 2001 | 11313958 |
| human itch is a coregulator of the hematopoietic transcription factor nf-e2. | we have cloned a new protein that interacts with the hematopoietic dna-binding transcription factor, p45/nf-e2, by screening a human erythroleukemia cell cdna library with the yeast two-hybrid approach. predicted peptide sequence and chromosomal mapping identified the cloned molecule to be the product of the human ortholog of the mouse itch gene, which has been implicated previously in the regulation of growth and differentiation of erythroid and lymphoid cells. transfection experiments indicate ... | 2001 | 11318614 |
| intersection of interferon and hypoxia signal transduction pathways in nitric oxide-induced tumor apoptosis. | activated macrophages play a central role in antitumor immunity. however, the stimuli that activate macrophages to kill tumor cells are not completely understood. because the center of solid tumors can be hypoxic, we hypothesized that hypoxia may be an important signal in activating macrophages to kill tumor cells. hypoxia stimulates ifn-primed macrophages to express the inducible nitric oxide synthase (nos2) and to synthesize nitric oxide (no). we show that this synergy between ifn and hypoxia ... | 2001 | 11325839 |
| cloning and identification of mypt3: a prenylatable myosin targetting subunit of protein phosphatase 1. | to identify novel protein phosphatase 1 (pp1)-interacting proteins, a yeast two-hybrid 3t3-l1 adipocyte cdna library was screened with the catalytic subunit of pp1 as bait. in the present work, the isolation, identification and initial biochemical characterization of a novel pp1-interacting protein, mypt3, which is homologous with the myosin phosphatase targetting subunit (mypt) family, is described. mypt3 aligns >99% with a region of mouse genomic dna clone rp23-156p23 and localizes to chromoso ... | 2001 | 11336659 |
| from xeroderma pigmentosum to the biological clock contributions of dirk bootsma to human genetics. | this paper commemorates the multiple contributions of dirk bootsma to human genetics. during a scientific 'bootsma' cruise on his sailing-boat 'de losbol', we visit a variety of scenery locations along the lakes and canals in friesland, passing the highlights of dirk bootsma's scientific oeuvre. departing from 'de fluessen', his homeport, with his phd work on the effect of x-rays and uv on cell cycle progression, we head for the pioneering endeavours of his team on mapping genes on human chromos ... | 2001 | 11341993 |
| identification of human and mouse hira-interacting protein-5 (hirip5), two mammalian representatives in a family of phylogenetically conserved proteins with a role in the biogenesis of fe/s proteins. | the human hira protein is encoded from a region of chromosome 22q that is critical for the digeorge syndrome and the velocardiofacial syndrome. we have previously reported that it directly interacts with core histones, with a novel histone-binding protein, hirip3, and during mouse embryogenesis, with the developmentally regulated homeodomain protein pax3, suggesting a promoter-targeted function at the chromatin level. we here report on hira-interacting protein 5 (hirip5), a small acidic protein ... | 2001 | 11342215 |
| effects of cysteine to serine substitutions in the two inter-chain disulfide bonds of insulin. | using site-directed mutagenesis we deleted the two inter-chain disulfide bonds of insulin, separately or both, by substitution of the cysteine residues with serine. deletion of a20-b19 or both of the two inter-chain disulfide bonds resulted in the complete loss of secretion of the mutant single-chain porcine insulin precursor (pip) from saccharomyces cerevisiae cells. removal of the a7-b7 disulfide bond resulted in a large reduction of secretion, but we could obtain the mutant for analysis of it ... | 2001 | 11347892 |
| friedreich's ataxia and frataxin: molecular genetics, evolution and pathogenesis (review). | friedreich's ataxia is an autosomal recessive neuro-degenerative disorder involving both central and peripheral nervous system. patients also show a systemic clinical picture presenting heart disease and diabetes mellitus or glucose intolerance. the disease is caused by mutations in the frda gene mapped on chromosome 9q13. the product of the gene is frataxin, an 18 kda soluble mitochondrial protein with 210 amino acids. crystal structure suggests a new, not previously reported, protein fold. the ... | 2001 | 11351269 |
| genomic organization, expression, and localization of murine ran-binding protein 2 (ranbp2) gene. | the ran-binding protein 2 (ranbp2) is a giant scaffold and mosaic cyclophilin-related nucleoporin implicated in the ran-gtpase cycle. there are no orthologs of the ranbp2 gene in yeast and drosophila genomes. in humans, this bona fide gene is partially duplicated in a ranbp2 gene cluster and lies in a hot spot for recombination on chromosome (chr) 2q. this genetic heterogeneity renders further significance of this genomic region in human disease due to its possible involvement in genetically lin ... | 2001 | 11353387 |
| unravelling the complex genetics of cleft lip in the mouse model. | nonsyndromic cleft lip in "a" strain mice and humans is genetically complex and is distinct from isolated cleft palate. cleft lip embryos recovered in 2.4% of 1485 first backcross (bc1) segregants from a cross of a/wysnj (24% cleft lip) and c57bl/6j (no cleft lip) in a/wysnj mothers, and in testcrosses of 10 recombinant inbred (ri) strains (axb/pgn or bxa/pgn), were used for gene mapping and for inference of genetic architecture. the a/wysnj maternal genotype increased cleft lip risk in reciproc ... | 2001 | 11353389 |
| dqx1, an rna-dependent atpase homolog with a novel deaq box: expression pattern and genomic sequence comparison of the human and mouse genes. | dqx1 is a novel gene related to the rna-dependent atpases. the gene was classified as a member of the dead/h family on the basis of the conserved order and spacing of ten short protein motifs. the unique features of dqx1 include replacement of the signature deah motif with deaq and the absence of the helicase motif. we determined the coding sequences of human and mouse dqx1, which encode proteins of 717 and 718 amino acids with 84% amino acid sequence identity. the 3.2-kb dqx1 transcript has hig ... | 2001 | 11353393 |
| identification and characterization of the cryptococcus neoformans phosphomannose isomerase-encoding gene, man1, and its impact on pathogenicity. | the polysaccharide capsule surrounding cryptococcus neoformans comprises manose, xylose and glucuronic acid, of which mannose is the major constituent. the gdp-mannose biosynthesis pathway is highly conserved in fungi and consists of three key enzymes: phosphomannose isomerase (pmi), phosphomannomutase (pmm) and gdp-mannose pyrophosphorylase (gmp). the man1 gene, encoding for the pmi enzyme, was isolated and sequenced from c. neoformans, and a disruption of the man1 gene was generated. one man1 ... | 2001 | 11359567 |
| ucs15a, a non-kinase inhibitor of src signal transduction. | src tyrosine kinase plays key roles in signal transduction following growth factor stimulation and integrin-mediated cell-substrate adhesion. since src-signal transduction defects are implicated in a multitude of human diseases, we have sought to develop new ways to identify small molecule inhibitors using a yeast-based, activated-src over-expression system. in the present study, we describe the identification of a unique src-signal transduction inhibitor, ucs15a. ucs15a was found to inhibit the ... | 2001 | 11360191 |
| regulation of the p21-activated kinase (pak) by a human gbeta -like wd-repeat protein, hpip1. | the family of p21-activated protein kinases (paks) is composed of serine-threonine kinases whose activity is regulated by the small guanosine triphosphatases (gtpases) rac and cdc42. in mammalian cells, paks have been implicated in the regulation of mitogen-activated protein cascades, cellular morphological and cytoskeletal changes, neurite outgrowth, and cell apoptosis. although the ability of cdc42 and rac gtpases to activate pak is well established, relatively little is known about the negati ... | 2001 | 11371639 |
| lung krüppel-like factor contains an autoinhibitory domain that regulates its transcriptional activation by binding wwp1, an e3 ubiquitin ligase. | lung krüppel-like factor (lklf/krüppel-like factor 2), a member of the krüppel-like factor family of transcription factors, is expressed predominantly in the lungs, with low levels of expression in other organs such as heart, spleen, skeletal muscle, and testis. lklf is essential during pulmonary development and single-positive t-cell development and is indispensable during mouse embryogenesis. in this study, we performed a series of experiments to define the activation domain of lklf as a means ... | 2001 | 11375995 |
| fibroblast growth factor homologous factor 1b binds to the c terminus of the tetrodotoxin-resistant sodium channel rnav1.9a (nan). | in this study we demonstrate a direct interaction between a cytosolic fibroblast growth factor family member and a sodium channel. a yeast two-hybrid screen for proteins that associate with the cytoplasmic domains of the tetrodotoxin-resistant sodium channel rna(v)1.9a (nan) led to the identification of fibroblast growth factor homologous factor 1b (fhf1b), a member of the fibroblast growth factor family, as an interacting partner of rna(v)1.9a. fhf1b selectively interacts with the c-terminal re ... | 2001 | 11376006 |
| three loci on mouse chromosome 6 influence onset and final incidence of type i diabetes in nod.c3h congenic strains. | the development of insulin-dependent diabetes mellitus in both human and mouse is dependent on the interaction between genetic and environmental factors. the analysis of newly created nod.c3h congenic strains for spontaneous and cyclophosphamide-induced diabetes has allowed the definition of three controlling genetic loci on mouse chromosome 6. a nod-derived susceptibility allele at the idd6 locus strongly influences the onset of diabetes in spontaneous diabetes. a nod-derived resistance allele ... | 2001 | 11386752 |
| mutations in cdh23 cause nonsyndromic hearing loss in waltzer mice. | mutations at the waltzer (v) locus result in deafness and vestibular dysfunction due to degeneration of the neuroepithelium within the inner ear. here, we use a positional cloning approach to show that waltzer encodes a novel cadherin (cdh23), which is most closely related to the drosophila fat protein. a single nucleotide deletion in the v(j) allele and a single nucleotide insertion in the v allele are predicted to truncate each protein near the n-terminus and produce a functional null allele. ... | 2001 | 11386759 |
| genetic and molecular characterization of skb15, a highly conserved inhibitor of the fission yeast pak, shk1. | the p21-activated kinase, shk1, is essential for viability, establishment and maintenance of cell polarity, and proper mating response in the fission yeast, schizosaccharomyces pombe. here we describe the characterization of a highly conserved, wd repeat protein, skb15, which negatively regulates shk1 in fission yeast. a null mutation in the skb15 gene is lethal and results in deregulation of actin polymerization and localization, microtubule biogenesis, and the cytokinetic machinery, as well as ... | 2001 | 11389855 |
| bcl-2 targets protein phosphatase 1 alpha to bad. | the diverse forms of protein phosphatase 1 (pp1) in vivo result from the association of the catalytic subunit with different regulatory subunits. we recently have described that pp1alpha is a ras-activated bad phosphatase that regulates il-2 deprivation-induced apoptosis. with the yeast two-hybrid system, gst fusion proteins, indirect immunofluorescence, and coimmunoprecipitation, we found that bcl-2 interacts with pp1alpha and bad. in contrast, bad did not interact with 14-3-3 protein. bcl-2 de ... | 2001 | 11390485 |
| rhythmic expression of nocturnin mrna in multiple tissues of the mouse. | nocturnin was originally identified by differential display as a circadian clock regulated gene with high expression at night in photoreceptors of the african clawed frog, xenopus laevis. although encoding a novel protein, the nocturnin cdna had strong sequence similarity with a c-terminal domain of the yeast transcription factor ccr4, and with mouse and human ests. since its original identification others have cloned mouse and human homologues of nocturnin/ccr4, and we have cloned a full-length ... | 2001 | 11394964 |
| recent molecular advances in studies of the concentrative na+-dependent nucleoside transporter (cnt) family: identification and characterization of novel human and mouse proteins (hcnt3 and mcnt3) broadly selective for purine and pyrimidine nucleosides (system cib). | the human concentrative (na+-linked) plasma membrane transport proteins hcnt1 and hcnt2, found primarily in specialized epithelia, are selective for pyrimidine nucleosides (system cit) and purine nucleosides (system cif), respectively. both have orthologs in other mammalian species and belong to a gene family (cnt) that also includes members in lower vertebrates, insects, nematodes, pathogenic yeast and bacteria. the cnt transporter family also includes a newly identified human and mouse cnt3 tr ... | 2001 | 11396613 |
| biochemical studies of zmpste24-deficient mice. | genetic studies in saccharomyces cerevisiae identified two genes, ste24 and rce1, involved in cleaving the three carboxyl-terminal amino acids from isoprenylated proteins that terminate with a caax sequence motif. ste24p cleaves the carboxyl-terminal "-aax" from the yeast mating pheromone a-factor, whereas rce1p cleaves the -aax from both a-factor and ras2p. ste24p also cleaves the amino terminus of a-factor. the mouse genome contains orthologues for both yeast rce1 and ste24. we previously demo ... | 2001 | 11399759 |
| mouse fatty acid transport protein 4 (fatp4): characterization of the gene and functional assessment as a very long chain acyl-coa synthetase. | fatp4 (slc27a4) is a member of the fatty acid transport protein (fatp) family, a group of evolutionarily conserved proteins that are involved in cellular uptake and metabolism of long and very long chain fatty acids. we cloned and characterized the murine fatp4 gene and its cdna. from database analysis we identified the human fatp4 genomic sequence. the fatp4 gene was assigned to mouse chromosome 2 band b, syntenic to the region 9q34 encompassing the human gene. the open reading frame was determ ... | 2001 | 11404000 |
| cloning, characterization and mapping of the mouse trehalase (treh) gene. | trehalase is the least studied of the membrane-bound alpha- glucosidase enzymes. here we report the isolation and characterization of the mouse trehalase (treh) gene. initially, pcr using primers based on published rat cdna sequence was used to clone a partial mouse cdna. this allowed design of mouse primers which identified a single positive clone in a bacterial artificial chromosome (bac) library of mouse genomic dna. analysis of bac subclones showed that the treh structural gene spans approxi ... | 2001 | 11404018 |
| heterogeneous rna-binding protein m4 is a receptor for carcinoembryonic antigen in kupffer cells. | here we report the isolation of the recombinant cdna clone from rat macrophages, kupffer cells (kc) that encodes a protein interacting with carcinoembryonic antigen (cea). to isolate and identify the cea receptor gene we used two approaches: screening of a kc cdna library with a specific antibody and the yeast two-hybrid system for protein interaction using as a bait the n-terminal part of the cea encoding the binding site. both techniques resulted in the identification of the rat heterogeneous ... | 2001 | 11406629 |
| requirement of cdc45 for postimplantation mouse development. | cdc45 is required for the initiation of dna replication in saccharomyces cerevisiae and functions as a dna polymerase alpha loading factor in xenopus, but its role in mammalian dna replication is unknown. to investigate the genetic and physiological functions of cdc45, we used a gene targeting strategy to generate mice lacking a functional cdc45 gene. homozygous mutant mice lacking a functional cdc45 gene underwent uterine implantation and induced uterine decidualization but did not develop subs ... | 2001 | 11416137 |
| the mammalian guanine nucleotide exchange factor msec12 is essential for activation of the sar1 gtpase directing endoplasmic reticulum export. | the sar1 gtpase is an essential component of copii vesicle coats involved in export of cargo from the endoplasmic reticulum of mammalian cells. to begin to elucidate its mechanism of action, we now report the identity of the mammalian homolog to the yeast sec12 guanine nucleotide exchange factor (18% identity) that promotes sar1 activation. mammalian sec12 (msec12) is a type ii transmembrane protein with a large cytosolic domain, a fragment of which has previously been reported as the transcript ... | 2001 | 11422940 |
| the synovial sarcoma associated protein syt interacts with the acute leukemia associated protein af10. | as a result of the synovial sarcoma associated t(x;18) translocation, the human syt gene on chromosome 18 is fused to either the ssx1 or the ssx2 gene on the x chromosome. although preliminary evidence indicates that the (fusion) proteins encoded by these genes may play a role in transcriptional regulation, little is known about their exact function. we set out to isolate interacting proteins through yeast two hybrid screening of a human cdna library using syt as a bait. of the positive clones i ... | 2001 | 11423977 |
| protein dispensability and rate of evolution. | if protein evolution is due in large part to slightly deleterious amino acid substitutions, then the rate of evolution should be greater in proteins that contribute less to individual fitness. the rationale for this prediction is that relatively dispensable proteins should be subject to weaker purifying selection, and should therefore accumulate mildly deleterious substitutions more rapidly. although this argument was presented over twenty years ago, and is fundamental to many applications of ev ... | 2001 | 11429604 |
| rim1 and rabphilin-3 bind rab3-gtp by composite determinants partially related through n-terminal alpha -helix motifs. | rim1 is a protein of the presynaptic active zone, the area of the plasma membrane specialized for neurotransmitter exocytosis, and interacts with rab3, a small gtpase implicated in neurotransmitter vesicle dynamics. here, we have studied the molecular determinants of rim1 that are responsible for rab3 binding, employing surface plasmon resonance and recombinant, bacterially expressed rab3 and rim1 proteins. a site that binds gtp- but not gdp-saturated rab3 was localized to a short alpha-helical ... | 2001 | 11431472 |
| molecular and biochemical characterization of rat epsilon -n-trimethyllysine hydroxylase, the first enzyme of carnitine biosynthesis. | epsilon-n-trimethyllysine hydroxylase (ec ) is the first enzyme in the biosynthetic pathway of l-carnitine and catalyzes the formation of beta-hydroxy-n-epsilon-trimethyllysine from epsilon-n-trimethyllysine, a reaction dependent on alpha-ketoglutarate, fe(2+), and oxygen. we purified the enzyme from rat kidney and sequenced two internal peptides by quadrupole-time-of-flight mass spectroscopy. the peptide sequences were used to search the expressed sequence tag data base, which led to the identi ... | 2001 | 11431483 |
| reconstruction of ligand-dependent transactivation of choristoneura fumiferana ecdysone receptor in yeast. | ecdysteroids play an important role in regulating development and reproduction in insects. interaction of 20-hydroxyecdysone (20e) with ecdysone receptor (ecr) as a heterodimer with ultraspiracle (usp) protein triggers the activation of 20e-responsive genes. in this paper we describe a ligand-mediated transactivation system in yeast using the spruce budworm choristoneura fumiferana ecdysone receptor (cfecr). coexpression of c. fumiferana usp (cfusp) with cfecr in yeast led to constitutive transc ... | 2001 | 11435614 |
| phosphorylation of mcm4 at specific sites by cyclin-dependent kinase leads to loss of mcm4,6,7 helicase activity. | mcm proteins that play an essential role in eukaryotic dna replication are phosphorylated in vivo, and cyclin-dependent protein kinase is at least in part responsible for the phosphorylation of mcm4. our group reported that the dna helicase activity of mcm4,6,7 complex, which may be involved in initiation of dna replication, is inhibited following phosphorylation by cdk2/cyclin a in vitro. here, we further examined the interplay between mouse mcm4,6,7 complex and cyclin-dependent kinases and det ... | 2001 | 11454864 |
| specific association of nitric oxide synthase-2 with rac isoforms in activated murine macrophages. | nitric oxide synthase-2 (nos2) is responsible for high-output nitric oxide production important in renal inflammation and injury. using a yeast two-hybrid assay, we identified rac2, a rho gtpase member, as a nos2-interacting protein. nos2 and rac2 proteins coimmunoprecipitated from activated raw 264.7 macrophages. the two proteins colocalized in an intracellular compartment of these cells. glutathione-s-transferase (gst) pull-down assays revealed that both rac1 and rac2 associated with gst-nos2 ... | 2001 | 11457725 |
| the mouse mps1p-like kinase regulates centrosome duplication. | the yeast mps1p protein kinase acts in centrosome duplication and the spindle assembly checkpoint. we demonstrate here that a mouse mps1p ortholog (esk, which we designate mmps1p) regulates centrosome duplication. endogenous mmps1p and overexpressed gfp-mmps1p localize to centrosomes and kinetochores in mouse cells. overexpression of gfp-mmps1p causes reduplication of centrosomes during s phase arrest. in contrast, a kinase-deficient mutant blocks centrosome duplication altogether. control of ce ... | 2001 | 11461705 |
| the orphan nuclear receptor tr2 interacts directly with both class i and class ii histone deacetylases. | a combination of in vivo and in vitro assays was employed to describe the ligand-independent interaction of the orphan nuclear receptor tr2 and histone deacetylase proteins. the repressive effect of tr2 on transcription of a luciferase reporter driven by a promoter containing a direct repeat-5 (dr5) derived from the human rarbeta gene was suppressed by the addition of the histone deacetylase inhibitor trichostatin a. immunoprecipitation with flag-epitope (mdykddddk)-tagged histone deacetylase pr ... | 2001 | 11463856 |
| induction of a mitosis delay and cell lysis by high-level secretion of mouse alpha-amylase from saccharomyces cerevisiae. | some foreign proteins are produced in yeast in a cell cycle-dependent manner, but the cause of the cell cycle dependency is unknown. in this study, we found that saccharomyces cerevisiae cells secreting high levels of mouse alpha-amylase have elongated buds and are delayed in cell cycle completion in mitosis. the delayed cell mitosis suggests that critical events during exit from mitosis might be disturbed. we found that the activities of pp2a (protein phosphatase 2a) and mpf (maturation-promoti ... | 2001 | 11472949 |
| mammalian golgi-associated bicaudal-d2 functions in the dynein-dynactin pathway by interacting with these complexes. | genetic analysis in drosophila suggests that bicaudal-d functions in an essential microtubule-based transport pathway, together with cytoplasmic dynein and dynactin. however, the molecular mechanism underlying interactions of these proteins has remained elusive. we show here that a mammalian homologue of bicaudal-d, bicd2, binds to the dynamitin subunit of dynactin. this interaction is confirmed by mass spectrometry, immunoprecipitation studies and in vitro binding assays. in interphase cells, b ... | 2001 | 11483508 |