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a tissue-specific coactivator of steroid receptors, identified in a functional genetic screen.steroid receptors mediate responses to lipophilic hormones in a tissue- and ligand-specific manner. to identify nonreceptor proteins that confer specificity or regulate steroid signaling, we screened a human cdna library in a steroid-responsive yeast strain. one of the identified cdnas, isolated in the screen as ligand effect modulator 6, showed no homology to yeast or caenorhabditis elegans proteins but high similarity to the recently described mouse coactivator pgc-1 and was accordingly termed ...200010713165
molecular characterization of kex1, a kexin-like protease in mouse pneumocystis carinii.expression screening of a pneumocystis carinii-infected mouse lung cdna library with specific monoclonal antibodies (mabs) led to the identification of a p. carinii cdna with extensive homology to subtilisin-like proteases, particularly fungal kexins and mammalian prohormone convertases. the 3.1 kb cdna contains a single open reading frame encoding 1011 amino acids. structural similarities to fungal kexins in the deduced primary amino acid sequence include a putative proenzyme domain delineated ...200010721706
yra1p, a conserved nuclear rna-binding protein, interacts directly with mex67p and is required for mrna export.mex67p and mtr2p constitute an essential mrna export complex that interacts with poly(a)+ rna and nuclear pore proteins. we have identified yra1p, an intranuclear protein with in vitro rna-rna annealing activity, which directly binds to mex67p. the complex between yra1p and mex67p was reconstituted in vitro and shown by uv-crosslinking to bind directly to rna. mutants of yra1 are impaired in nuclear poly(a)+ rna export at restrictive growth conditions. aly, the mouse homologue of yra1p and a tra ...200010722314
comparison of nucleosome remodeling by the yeast transcription factor pho4 and the glucocorticoid receptor.chromatin reorganization of the pho5 and murine mammary tumor virus (mmtv) promoters is triggered by binding of either pho4 or the glucocorticoid receptor (gr), respectively. in order to compare the ability of pho4 and gr to remodel chromatin and activate transcription, hybrid promoter constructs were created by insertion of the mmtv b nucleosome sequence into the pho5 promoter and then transformed into a yeast strain expressing gr. activation of either pho4 (by phosphate depletion) or gr (by ho ...200010722753
a novel nuclear phosphoprotein, ganp, is up-regulated in centrocytes of the germinal center and associated with mcm3, a protein essential for dna replication.antigen (ag) immunization induces formation of the germinal center (gc), with large, rapidly proliferating centroblasts in the dark zone, and small, nondividing centrocytes in the light zone. we identified a novel nuclear protein, ganp, that is up-regulated in centrocytes. we found that ganp was up-regulated in gc b cells of peyer's patches in normal mice and in spleens from ag-immunized mice. ganp-positive cells appeared in the light zone of the gc, with coexpression of the peanut agglutinin (p ...200010733502
the survival motor neuron protein interacts with the transactivator fuse binding protein from human fetal brain.to identify interacting proteins of survival motor neuron (smn) in neurons, a fetal human brain cdna library was screened using the yeast two-hybrid system. one identified group of smn interacting clones encoded the dna transactivator fuse binding protein (fbp). fbp overexpressed in hek293 cells or endogenously expressed in fetal and adult mouse brain bound specifically in vitro to recombinant smn protein. furthermore, an anti-fbp antibody specifically co-immunoprecipitated smn when both protein ...200010734235
rab geranylgeranyl transferase alpha mutation in the gunmetal mouse reduces rab prenylation and platelet synthesis.few molecular events important to platelet biogenesis have been identified. mice homozygous for the spontaneous, recessive mutation gunmetal (gm) have prolonged bleeding, thrombocytopenia, and reduced platelet alpha- and delta-granule contents. here we show by positional cloning that gm results from a g-->a substitution mutation in a splice acceptor site within the alpha-subunit of rab geranylgeranyl transferase (rabggta), an enzyme that attaches geranylgeranyl groups to rab proteins. most rabgg ...200010737774
interaction of a novel cysteine and histidine-rich cytoplasmic protein with galectin-3 in a carbohydrate-independent manner.we have used the yeast two-hybrid system to search for cytoplasmic proteins that might assist in the intracellular trafficking of the soluble beta-galactoside-binding protein, galectin-3. we utilised as bait murine full-length galectin-3 to screen a murine 3t3 cdna library. several interacting clones were found to encode a partial open reading frame and a full-length clone was obtained by rapid amplification of cdna ends methodology. in various assays in vitro the novel protein was shown to bind ...200010745073
functional interaction between the mouse notch1 intracellular region and histone acetyltransferases pcaf and gcn5.the notch receptor that plays an important role in cell fate determination is intracellularly cleaved by interaction with the ligand. the cleaved intracellular region (ramic) of notch is translocated into the nucleus and interacts with a dna-binding protein rbp-j to activate transcription of genes that regulate cell differentiation. although ramic has been shown to facilitate the rbp-j-mediated transactivation by displacing the histone deacetylase corepressor complex from rbp-j, there is no evid ...200010747963
both normal and transforming pcph proteins have guanosine diphosphatase activity but only the oncoprotein cooperates with ras in activating extracellular signal-regulated kinase erk1.previous reports from our laboratory described the activation of the pcph gene into the pcph oncogene (mt-pcph, reported previously as cph) by a single point mutational deletion. as a consequence, the mt-pcph oncoprotein is a truncated form of the normal pcph protein. although both proteins have ribonucleotide diphosphate-binding activity, only mt-pcph acted synergistically with a human h-ras oncoprotein to transform murine nih3t3 fibroblasts. we report here the expression of the pcph and mt-pcp ...200010749145
the mouse neurological mutant flailer expresses a novel hybrid gene derived by exon shuffling between gnb5 and myo5a.exon shuffling is thought to be an important mechanism for evolution of new genes. here we show that the mouse neurological mutation flailer (flr) expresses a novel gene that combines the promoter and first two exons of guanine nucleotide binding protein beta 5 (gnb5) with the c-terminal exons of the closely linked myosin 5a (myova) gene (myo5a). the flailer protein, which is expressed predominantly in brain, contains the n-terminal 83 amino acids of gnb5 fused in-frame with the c-terminal 711 a ...200010749990
identification and characterization of a novel protein from sertoli cells, pass1, that associates with mammalian small stress protein hsp27.hsp27 is involved in development of tolerance to stress, possibly by its involvement in molecular chaperoning, maintenance of glutathione status, and/or modulation of microfilament structure and function. we hypothesize that hsp27 function depends on specific association with other proteins. to discover proteins that associate with hsp27, we made a differentiated rat sertoli cell cdna expression library and screened it using the yeast two-hybrid system. we obtained a cdna coding for a novel prot ...200010751411
drg represents a family of two closely related gtp-binding proteins.in a previous publication we identified a novel human gtp-binding protein that was related to drg, a developmentally regulated gtp-binding protein from the central nervous system of mouse. here we demonstrate that both the human and the mouse genome possess two closely related drg genes, termed drg1 and drg2. the two genes share 62% sequence identity at the nucleotide and 58% identity at the protein level. the corresponding proteins appear to constitute a separate family within the superfamily o ...200010760581
alternative splicing of murine sorcs leads to two forms of the receptor that differ completely in their cytoplasmic tails.we report the identification of a splice variant of sorcs, a member of the family of vps10 domain containing receptors. these type i transmembrane proteins share the presence of internalization signals in their cytoplasmic tail as one common characteristic. we show that an alternatively spliced transcript of sorcs is generated by differential processing of a composite internal/terminal exon. this splice variant encodes a protein with an n-terminal vps10 domain followed by a leucine-rich module a ...200010760602
the interaction between een and abi-1, two mll fusion partners, and synaptojanin and dynamin: implications for leukaemogenesis.the mixed lineage leukaemia gene, mll (also called hrx, all-1) in acute leukaemia is fused to at least 16 identified partner genes that display diverse structural and biochemical properties. using gst pull down and the yeast two hybrid system, we show that two different mll fusion partners with sh3 domains, een and abi-1, interact with dynamin and synaptojanin, both of which are involved in endocytosis. synaptojanin, a member of the inositol phosphatase family that has recently been shown to reg ...200010764144
specific sequence of motifs of mitochondrial uncoupling proteins.we have searched for the exclusivity of common sequence motifs of the mitochondrial uncoupling proteins (ucp1, ucp2, ucp3, ucp4, bmcp1, and plant ucp [pump]) within the gene family of mitochondrial anion carrier proteins. the ucp-specific sequences, "ucp signatures", were found in the first, second, and fourth alpha-helices. first: ala/ser-cys/thr/n-n/phe-ala/gly-[negatively charged residue]-n/phe-n/cys-thr-phe/n; second: gly/ala-ile/leu-gln/x-[positively charged residue]-nh-n/cys-ser/nphi/x-n/s ...200010772343
a sequence-specific rna binding complex expressed in murine germ cells contains msy2 and msy4.the protamine mrnas are stored for up to 8 days as translationally repressed ribonucleoprotein particles during murine spermatogenesis. translational repression of the protamine 1, prm1, mrna is controlled by sequences in its 3'-untranslated region (utr). in this study we used the yeast three-hybrid system to clone msy4, which encodes a novel member of the y box family of nucleic acid binding proteins. msy4 specifically binds to a site within the 5' most 37 nucleotides in the prm1 3' utr. msy4 i ...200010772793
mouse linkage cytogenetics (l-c) probes.we have identified 149 hybridization probes at 10-cm intervals in the mouse and have confirmed their order and linkage by fluorescence in situ hybridization. these probes represent a new resource for mapping in the mouse and can be used to correlate linkage and cytogenetic maps, to map novel sequences to within a few centimorgans, to relate cytogenetic abnormalities to the genetic map, and to make cross-species comparisons.200010773692
transcriptional activation by hepatocyte nuclear factor-1 requires synergism between multiple coactivator proteins.hepatocyte nuclear factor-1 (hnf-1) plays an important role in the regulation of a large number of genes expressed in the liver, kidney, and pancreatic beta-cells. in exploring the molecular mechanism involved in hnf-1-dependent gene activation in the in vivo chromatin context, we found that hnf-1 can physically interact with the histone acetyltransferases (hats) creb-binding protein (cbp), p300/cbp-associated factor (p/caf), src-1, and rac3. the transcriptional activation potential of hnf-1 on ...200010777539
cloning and expression analysis of a novel gene, rp42, mapping to an autism susceptibility locus on 6q16.we isolated a novel mouse gene, rp42, in a systematic search for genes expressed in proliferating neuroblasts whose human orthologs map to susceptibility loci for autism. this gene is intronless and encodes a putative 259-amino-acid protein that exhibits 30-36% overall sequence identity to a fission yeast and a nematode protein (genpept accession nos. caa17006 and cab54261). nevertheless, no homology to any known gene was found. rp42 has developmentally regulated expression, particularly in prol ...200010777668
faithful expression of the human 5q31 cytokine cluster in transgenic mice.interleukins -4, -5, and -13, cardinal cytokines produced by th2 cells, are coordinately expressed and clustered in 150-kb syntenic regions on mouse chromosome 11 and human chromosome 5q31. we analyzed two sets of human yeast artificial chromosome transgenic mice that contained the 5q31 cytokines to assess whether conserved sequences required for their coordinate and cell-specific regulation are contained within the cytokine cluster itself. human il-4, il-13, and il-5 were expressed under th2, b ...200010779759
ref, an evolutionary conserved family of hnrnp-like proteins, interacts with tap/mex67p and participates in mrna nuclear export.vertebrate tap and its yeast ortholog mex67p are involved in the export of messenger rnas from the nucleus. tap has also been implicated in the export of simian type d viral rnas bearing the constitutive transport element (cte). although tap directly interacts with cte-bearing rnas, the mode of interaction of tap/mex67p with cellular mrnas is different from that with the cte rna and is likely to be mediated by protein-protein interactions. here we show that mex67p directly interacts with yra1p, ...200010786854
isolation and characterization of peroxisome proliferator-activated receptor (ppar) interacting protein (prip) as a coactivator for ppar.we previously isolated and identified steroid receptor coactivator-1 (src-1) and peroxisome proliferator-activated receptor (ppar)-binding protein (pbp/pparbp) as coactivators for ppar, using the ligand-binding domain of ppargamma as bait in a yeast two-hybrid screening. as part of our continuing effort to identify cofactors that influence the transcriptional activity of ppars, we now report the isolation of a novel coactivator from mouse, designated prip (peroxisome proliferator-activated recep ...200010788465
binding of adam12, a marker of skeletal muscle regeneration, to the muscle-specific actin-binding protein, alpha -actinin-2, is required for myoblast fusion.adam12 belongs to the transmembrane metalloprotease adam ("a disintegrin and metalloprotease") family. adam12 has been implicated in muscle cell differentiation and fusion, but its precise function remains unknown. here, we show that adam12 is dramatically up-regulated in regenerated, newly formed fibers in vivo. in c2c12 cells, adam12 is expressed at low levels in undifferentiated myoblasts and is transiently up-regulated at the onset of differentiation when myoblasts fuse into multinucleated m ...200010788519
rcl1p, the yeast protein similar to the rna 3'-phosphate cyclase, associates with u3 snornp and is required for 18s rrna biogenesis.rna 3'-terminal phosphate cyclases are evolutionarily conserved enzymes catalysing conversion of the 3'-terminal phosphate in rna to the 2',3'-cyclic phosphodiester. their biological role remains unknown. the yeast saccharomyces cerevisiae contains a gene encoding a protein with strong sequence similarity to the characterized cyclases from humans and escherichia coli. the gene, named rcl1 (for rna terminal phosphate cyclase like), is essential for growth, and its product, rcl1p, is localized in ...200010790377
structural analysis of ww domains and design of a ww prototype.two new nmr structures of ww domains, the mouse formin binding protein and a putative 84.5 kda protein from saccharomyces cerevisiae, show that this domain, only 35 amino acids in length, defines the smallest monomeric triple-stranded antiparallel beta-sheet protein domain that is stable in the absence of disulfide bonds, tightly bound ions or ligands. the structural roles of conserved residues have been studied using site-directed mutagenesis of both wild type domains. crucial interactions resp ...200010802733
the roles of the mcm, orc, and cdc6 proteins in determining the replication competence of chromatin in quiescent cells.most eukaryotic cell types can withdraw from proliferative cell cycles and remain quiescent for extended periods. intact nuclei isolated from quiescent murine nih3t3 cells fail to replicate in vitro when incubated in xenopus egg extracts, although intact nuclei from proliferating cells replicate well. permeabilization of the nuclear envelope rescues the ability of quiescent nuclei to replicate in the extract. we show that origin replication complex (orc), minichromosome maintenance (mcm), and cd ...200010806069
molecular cloning and expression analysis of the human da41 gene and its mapping to chromosome 9q21.2-q21.3.da41 was previously identified as one of the dan-binding proteins, via a yeast-based two-hybrid screening strategy. in the present study, we cloned a human homolog of da41 cdna. structural analysis revealed that human da41 cdna consisted of 2,861 nucleotides in length and encoded a protein of 589 amino acids, with a predicted molecular mass of 62.4kda. human da41 exhibited an 86% amino acid sequence identity to rat da41, indicating the evolutionarily conserved structure and function of da41. a d ...200010807547
cell type-dependent transactivation or repression of mesoderm-restricted basic helix-loop-helix protein, pod-1/capsulin.a family of basic-helix-loop-helix (bhlh) nuclear factors play important roles in controlling cell growth and differentiation as critical regulatory components in transcription. here we describe molecular characterization of mesoderm-specific bhlh protein, pod-1/capsulin. transactivation property of pod-1/capsulin was analyzed by the gal4 fusion system in six mammalian cell lines. the results indicated that an activation property was shown in ht1080 and hela cells, but a repression activity in h ...200010821432
querkopf, a myst family histone acetyltransferase, is required for normal cerebral cortex development.in order to find, and mutate, novel genes required for regulation of neurogenesis in the cerebral cortex, we performed a genetic screen in mice. as the result of this screen, we created a new mouse mutant, querkopf. the querkopf mutation is due to an insertion into a myst family histone acetyltransferase gene. mice homozygous for the querkopf mutation have craniofacial abnormalities, fail to thrive in the postnatal period and have defects in central nervous system development. the defects in cen ...200010821753
induction of human cdc37 in prostate cancer correlates with the ability of targeted cdc37 expression to promote prostatic hyperplasia.the cdc37 gene encodes a 50 kda protein which targets intrinsically unstable oncoprotein kinases such as cdk4, raf-1, and src to the molecular chaperone hsp90. this activity is thought to play an important role in the establishment of signaling pathways controlling cell proliferation. the budding yeast cdc37 homolog is required for cell division and mammalian cdc37 is expressed in proliferative zones during embryonic development and in adult tissues, consistent with a positive role in proliferat ...200010822368
the oncoprotein kinase chaperone cdc37 functions as an oncogene in mice and collaborates with both c-myc and cyclin d1 in transformation of multiple tissues.cdc37 encodes a 50-kda protein that targets intrinsically unstable oncoprotein kinases including cdk4, raf-1, and v-src to the molecular chaperone hsp90, an interaction that is thought to be important for the establishment of signaling pathways. cdc37 is required for proliferation in budding yeast and is coexpressed with cyclin d1 in proliferative zones during mouse development, a finding consistent with a positive role in cell proliferation. cdc37 expression may not only be required to support ...200010825210
hrs interacts with snap-25 and regulates ca(2+)-dependent exocytosis.synaptosome-associated protein of 25 kda (snap-25) is a neuronal membrane protein essential for synaptic vesicle exocytosis. to investigate the mechanisms by which snap-25 mediates neurosecretion, we performed a search for proteins that interact with snap-25 using a yeast two-hybrid screen. here, we report the isolation and characterization of a snap-25-interacting protein that is the rat homologue of mouse hepatocyte growth factor-regulated tyrosine kinase substrate (hrs). hrs specifically inte ...200010825299
the candida albicans antiporter gene cnh1 has a role in na+ and h+ transport, salt tolerance, and morphogenesis.the isolation and functional characterization of a candida albicans na+/h+ antiporter gene, cnh1, is reported here. the gene encodes a protein of 840 amino acids that exhibits high levels of similarity in sequence, size, and structural and functional domains to a group of known na+/h+ antiporters of fungi. the cnh1 gene is able to functionally complement the salt-sensitivity of a saccharomyces cerevisiae ena1 nha1 mutant, and mutations of two conserved aspartate residues to asparagines in the pu ...200010832630
selection of homeotic proteins for binding to a human dna replication origin.we have previously shown that a cell cycle-dependent nucleoprotein complex assembles in vivo on a 74 bp sequence within the human dna replication origin associated to the lamin b2 gene. here, we report the identification, using a one-hybrid screen in yeast, of three proteins interacting with the 74 bp sequence. all of them, namely hoxa13, hoxc10 and hoxc13, are orthologues of the abdominal-b gene of drosophila melanogaster and are members of the homeogene family of developmental regulators. we d ...200010835276
the nuclear dead box rna helicase p68 interacts with the nucleolar protein fibrillarin and colocalizes specifically in nascent nucleoli during telophase.the dead box protein, p68, is an established rna-dependent atpase and rna helicase in vitro, but neither the physiological function of this protein nor the macromolecules with which it interacts are known. using a yeast two-hybrid screen, we identified the nucleolar protein, fibrillarin, as a protein that interacts with p68. coimmunoprecipitation experiments confirmed that p68 and fibrillarin can form complexes in cellular extracts, and deletion analysis identified regions in each protein respon ...200010837141
molecular characterization of human acetyl-coa synthetase, an enzyme regulated by sterol regulatory element-binding proteins.through suppressive subtractive hybridization, we identified a new gene whose transcription is induced by sterol regulatory element-binding proteins (srebps). the gene encodes acetyl-coa synthetase (acs), the cytosolic enzyme that activates acetate so that it can be used for lipid synthesis or for energy generation. acs genes were isolated previously from yeast, but not from animal cells. recombinant human acs was produced by expressing the cloned cdna transiently in human cells. after purificat ...200010843999
interaction between the transcription factor spbp and the positive cofactor rnf4. an interplay between protein binding zinc fingers.the activator of stromelysin 1 gene transcription, spbp, interacts with the ring finger protein rnf4. both proteins are ubiquitously expressed and localized in the nucleus. rnf4 facilitates accumulation of specific spbp-dna complexes in vitro and acts as a positive cofactor in spbp-mediated transactivation. spbp harbors an internal zinc finger of the phd/lap type. this domain can form intra-chain protein-protein contacts in spbp resulting in negative modulation of spbp-rnf4 interaction.200010849425
identification and characterization of an spo11 homolog in the mouse.the spo11/topvia family includes proteins from archaebacteria and eukaryotes. the protein member from the archaebacterium sulfulobus shibatae is the catalytic subunit of topovi dna topoisomerase. in saccharomyces cerevisiae, schizosaccharomyces pombe, caenorhabditis elegans and drosophila melanogaster, spo11 is required for meiotic recombination, suggesting a conserved mechanism for the initiation step of this process. indeed, s. cerevisiae spo11 has been shown to be directly involved in the for ...200010855504
cloning and characterization of harp/smarcal1: a prokaryotic hepa-related snf2 helicase protein from human and mouse.the snf2 gene family consists of a large group of proteins involved in transcriptional regulation, maintenance of chromosome integrity, and various aspects of dna repair. we cloned a novel snf2 family human cdna, with sequence identity to the escherichia coli rna polymerase-binding protein hepa and named the human hepa-related protein (hharp/smarcal1). in addition, the mouse ortholog (mharp/smarcal1) was cloned, and the caenorhabditis elegans ortholog (ceharp) was identified in the genbank datab ...200010857751
functional analysis of a tryptophan-less p-glycoprotein: a tool for tryptophan insertion and fluorescence spectroscopy.p-glycoprotein (pgp) functions as an atp-dependent drug efflux pump to confer multidrug resistance to tumor cells. in the absence of a high-resolution structure for this protein, several important and intriguing aspects of pgp structure and function remain poorly understood. fluorescence spectroscopy of endogenous or genetically engineered tryptophan residues represents a potentially powerful method to probe static and dynamic aspects of pgp at high resolution. we have used site-directed mutagen ...200010860925
identification of four single nucleotide polymorphisms in dna repair genes: xpa and xpb (ercc3) in polish population.a deficiency in dna repair is associated with increased cancer risk. inter-individual variations in dna repair capacity observed in humans may result from genetic polymorphisms in dna repair genes. in order to provide a basis for future functional and molecular epidemiology studies on cancer susceptibility, we screened 35 individuals for polymorphisms in coding regions of xpa and xpb genes involved in nucleotide excision repair (ner). relevant cdna sequences were amplified by pcr, sequenced with ...200010862089
altered expression of fatty acid-metabolizing enzymes in aromatase-deficient mice.hepatic steatosis is a frequent complication in nonobese patients with breast cancer treated with tamoxifen, a potent antagonist of estrogen. in addition, hepatic steatosis became evident spontaneously in the aromatase-deficient (arko) mouse, which lacks intrinsic estrogen production. these clinical and laboratory observations suggest that estrogen helps to maintain constitutive lipid metabolism. to clarify this hypothesis, we characterized the expression and activity in arko mouse liver of enzy ...200010862797
human sphingosine kinase: molecular cloning, functional characterization and tissue distribution.sphingosine-1-phosphate (spp), the product of sphingosine kinase, is an important signaling molecule with intra- and extracellular functions. the cdna for the mouse sphingosine kinase has recently been reported. in this paper we describe the cloning, expression and characterization of the human sphingosine kinase (husphk1). sequence analysis comparison revealed that this kinase is evolutionarily very conserved, having a high degree of homology with the murine enzyme, and presenting several conse ...200010863092
box c/d snorna-associated proteins: two pairs of evolutionarily ancient proteins and possible links to replication and transcription.the eukaryotic nucleolus contains a diverse population of small nucleolar rnas (snornas) essential for ribosome biogenesis. the box c/d snorna family possesses conserved nucleotide boxes c and d that are multifunctional elements required for snorna processing, snorna transport to the nucleolus, and 2'-o-methylation of ribosomal rna. we have previously demonstrated that the assembly of an snornp complex is essential for processing the intronic box c/d snornas and that specific nuclear proteins as ...200010864044
yeast tissue phase of emmonsia pasteuriana inoculated in golden hamster by intratesticular way.the scope of our study was to present an experimental model reproducing the dimorphic yeast-like population (as for histoplasma capsulatum, blastomyces dermatitidis) similar to that observed in the cutaneous biopsy of an italian woman who had never traveled abroad, being intravenous drug user and hiv positive for 10 years, finally infected with the new dimorphic fungus emmonsia pasteuriana. experimental inoculation was unsuccessful by intraperitoneal (i.p.) and intravenous (i.v.) ways in a mouse ...199910865897
identification of a novel beta-catenin-interacting protein.cadherin is a well-known cell-cell adhesion molecule, and it binds to beta-catenin, which in turn binds to alpha-catenin. however, little is known about the regulatory mechanism underlying the cadherin-mediated cell-cell adhesion. here we purified two novel beta-catenin-interacting proteins with molecular masses of 180 kda (p180) and 150 kda (p150) from bovine brain cytosol by using glutathione s-transferase (gst)-beta-catenin affinity column chromatography. mass spectral analysis revealed p180 ...200010873669
reversal of fortune for drosophila geneticists?although drosophila is a wonderful model organism, there is one molecular arena where it lags far behind its yeast and mouse model counterparts. reverse genetics, whereby a piece of dna is integrated into a target gene such that the gene is disrupted or replaced, is not easy in drosophila. as engels explains in his provocative perspective, this may be set to change with the description of a new method for reverse genetics in drosophila (rong and golic). this new technique should ensure that dros ...200010877715
ariadne-1: a vital drosophila gene is required in development and defines a new conserved family of ring-finger proteins.we report the identification and functional characterization of ariadne-1 (ari-1), a novel and vital drosophila gene required for the correct differentiation of most cell types in the adult organism. also, we identify a sequence-related gene, ari-2, and the corresponding mouse and human homologues of both genes. all these sequences define a new protein family by the acid-rich, ring finger, b-box, ring finger, coiled-coil (arbrcc) motif string. in drosophila, ari-1 is expressed throughout develop ...200010880484
involvement of the trap220 component of the trap/smcc coactivator complex in embryonic development and thyroid hormone action.the trap220 component of the trap/smcc complex, a mammalian homologof the yeast mediator that shows diverse coactivation functions, interacts directly with nuclear receptors. ablation of the murine trap220 gene revealed that null mutants die during an early gestational stage with heart failure and exhibit impaired neuronal development with extensive apoptosis. primary embryonic fibroblasts derived from null mutants show an impaired cell cycle regulation and a prominent decrease of thyroid hormon ...200010882104
electron microscopic observation and single-stranded dna binding activity of the mcm4,6,7 complex.mcm2-7 proteins that play an essential role in eukaryotic dna replication contain dna-dependent atpase motifs in a central domain that, from yeast to mammals, is highly conserved. our group has reported that a dna helicase activity is associated with a 600 kda human mcm4, 6 and 7 complex. the structure of the mcm4,6,7 complex was visualized by electron microscopy after negative staining with uranyl acetate. the complex contained toroidal forms with a central channel and also contained structures ...200010884341
cloning and expression of a cdna coding for catalase from zebrafish (danio rerio).a full-length complementary dna (cdna) clone encoding a catalase was amplified by the rapid amplication of cdna ends-polymerase chain reaction (race-pcr) technique from zebrafish (danio rerio) mrna. nucleotide sequence analysis of this cdna clone revealed that it comprised a complete open reading frame coding for 526 amino acid residues and that it had a molecular mass of 59 654 da. the deduced amino acid sequence showed high similarity with the sequences of catalase from swine (86.9%), mouse (8 ...200010888504
overexpression of murine fizzy-related (fzr) increases natural killer cell-mediated cell death and suppresses tumor growth.fizzy-related (fzr) is a recently identified 7wd domain family member implicated in cell cycle regulation of drosophila and yeast. in this study, the murine homologue of fzr was isolated by suppression subtractive hybridization as a gene with decreased expression during malignant progression of a murine b-lymphoma cell line. retroviral overexpression of fzr in b-lymphoma cells reduced tumor formation. those tumors that did arise had diminished or extinguished retroviral fzr. surprisingly, fzr ov ...200010891459
transcriptional regulation of the mouse cytosolic chaperonin subunit gene ccta/t-complex polypeptide 1 by selenocysteine trna gene transcription activating factor family zinc finger proteins.the chaperonin containing t-complex polypeptide 1 (cct) is a molecular chaperone assisting in the folding of proteins in eukaryotic cytosol, and the ccta (encoding the alpha subunit of cct)/t-complex polypeptide 1 gene encodes the alpha subunit of cct. we show here that transcription of the mouse ccta gene is regulated by selenocysteine trna gene transcription activating factor (staf) family zinc-finger transcription factors znf143 and znf76. reporter gene assay using hela cells indicated that t ...200010893243
assignment of the proline-rich protein gene prp2 (alias d6mit13) to mouse chromosome 6g1 by in situ hybridization. 200010894928
cdc42 is required for pip(2)-induced actin polymerization and early development but not for cell viability.cdc42 and other rho gtpases are conserved from yeast to humans and are thought to regulate multiple cellular functions by inducing coordinated changes in actin reorganization and by activating signaling pathways leading to specific gene expression. direct evidence implicating upstream signals and components that regulate cdc42 activity or for required roles of cdc42 in activation of downstream protein kinase signaling cascades is minimal, however. also, whereas genetic analyses have shown that c ...200010898977
hash-1 and e2-2 are expressed in human neuroblastoma cells and form a functional complex.the basic helix-loop-helix (bhlh) transcription factor mammalian achaete-scute homolog-1 (mash-1 in mouse and hash-1 in human) is essential for proper development of olfactory and most peripheral autonomic neurons, and for the formation of distinct neuronal circuits within the central nervous system. we have previously shown that hash-1 is expressed in neuroblastoma tumors and cell lines, and in this study we have used the yeast two-hybrid system to isolate hash-1 interacting proteins from a hum ...200010903890
the alpha subunits of gz and gi interact with the eyes absent transcription cofactor eya2, preventing its interaction with the six class of homeodomain-containing proteins.yeast two-hybrid techniques were used to identify possible effectors for the heterotrimeric g protein g(z) in human bone marrow cells. eya2, a human homologue of the drosophila eya transcription co-activator, was identified. eya2 interacts with activated galpha(z) and at least one other member of the galpha(i) family, galpha(i2). interactions were confirmed in mammalian two-hybrid and glutathione s-transferase fusion protein pull-down assays. regions of eya2-mediating interaction were mapped to ...200010906137
molecular characterization of a granulocyte macrophage-colony-stimulating factor receptor alpha subunit-associated protein, grap.the granulocyte macrophage-colony-stimulating factor receptor (gm-csf-r) is a heterodimer composed of 2 subunits, alpha and beta, and ligand binding to the high-affinity receptor leads to signalling for the multiple actions of gm-csf on target cells. in order to explore the role of the alpha subunit in signalling, we used a yeast-2-hybrid system to identify proteins interacting with the intracellular domain of the gmr-alpha. a cdna encoding a predicted protein of 198 amino acids, designated grap ...200010910889
in vivo interaction of the adapter protein cd2-associated protein with the type 2 polycystic kidney disease protein, polycystin-2.we identified a developmentally regulated gene from mouse kidney whose expression is up-regulated in metanephrogenic mesenchyme cells when they are induced to differentiate to epithelial cells during kidney organogenesis. the deduced 70.5-kda protein, originally named mets-1 (mesenchyme-to-epithelium transition protein with sh3 domains), has since been cloned as a cd2-associated protein (cd2ap). cd2ap is strongly expressed in glomerular podocytes, and the absence of cd2ap in mice results in cong ...200010913159
inactivation of mouse hus1 results in genomic instability and impaired responses to genotoxic stress.the eukaryotic cell cycle is overseen by regulatory mechanisms, termed checkpoints, that respond to dna damage, mitotic spindle defects, and errors in the ordering of cell cycle events. the dna replication and dna damage cell cycle checkpoints of the fission yeast schizosaccharomyces pombe require the hus1(+) (hydroxyurea sensitive) gene. to determine the role of the mouse homolog of hus1(+) in murine development and cell cycle checkpoint function, we produced a targeted disruption of mouse hus1 ...200010921903
sequence diversification of the fk506-binding proteins in several different genomes.sequences of fk506-binding proteins (fkbps) from four genomes of the following organisms were compared: the prokaryote escherichia coli, the lower eukaryote saccharomyces cerevisiae, the plant arabidopsis thaliana, the nematode caenorhabditis elegans and a composite of 14 unique fkbps from two mammalian organisms homo sapiens (man) and mus musculus (domestic mouse). a singular fk506-like binding domain (fkbd) has about 12 kda and occurs in the form of archetypal fkbp-12 and as a part of differen ...200010931176
a novel method for constructing gene-targeting vectors.we developed a simple and rapid method for constructing knockout vectors using inverse-pcr (ipcr). the method consists of three steps: (i) digestion of a target bacterial artificial chromosome with several restriction enzymes (six-base cutters) followed by self-ligation; (ii) ipcr using circular dnas as templates and two primers which are oriented in opposite directions; and (iii) cloning into a vector containing a positive selection marker, which results in a typical replacement knockout vector ...200010931936
creb-binding protein/p300 activates myod by acetylation.the myogenic protein myod requires two nuclear histone acetyltransferases, creb-binding protein (cbp)/p300 and pcaf, to transactivate muscle promoters. myod is acetylated by pcaf in vitro, which seems to increase its affinity for dna. we here show that myod is constitutively acetylated in muscle cells. in vitro, myod is acetylated both by cbp/p300 and by pcaf on two lysines located at the boundary of the dna binding domain. myod acetylation by cbp/p300 (as well as by pcaf) increases its activity ...200010944526
prevalence and distribution of ret/ptc 1, 2, and 3 in papillary thyroid carcinoma in new caledonia and australia.the world's highest incidence of thyroid cancer has been reported among females in new caledonia, a french overseas territory in the pacific located between australia and fiji. to date, no molecular genetic studies in this population are available. over the past few years, the oncogenic rearrangement of the ret protooncogene (ret/ptc) has been studied in papillary carcinomas in different populations. in this study, we investigated the prevalence and distribution of ret/ptc1, 2, and 3 in papillar ...200010946873
aven, a novel inhibitor of caspase activation, binds bcl-xl and apaf-1.bcl-x(l), an antiapoptotic bcl-2 family member, is postulated to function at multiple stages in the cell death pathway. the possibility that bcl-x(l) inhibits cell death at a late (postmitochondrial) step in the death pathway is supported by this report of a novel apoptosis inhibitor, aven, which binds to both bcl-x(l) and the caspase regulator, apaf-1. identified in a yeast two-hybrid screen, aven is broadly expressed and is conserved in other mammalian species. only those mutants of bcl-x(l)th ...200010949025
the adenomatous polyposis coli-protein (apc) interacts with the protein tyrosine phosphatase ptp-bl via an alternatively spliced pdz domain.mutations of the tumor suppressor protein apc (adenomatous polyposis coli) are linked to familiar and sporadic human colon cancer. here we describe a novel interaction between the apc protein and the protein tyrosine phosphatase ptp-bl carrying five pdz protein-protein interaction domains. exclusively, the second pdz domain (pdz2) of ptp-bl is binding to the extreme c-terminus of the apc protein, as determined by yeast two-hybrid studies. using surface plasmon resonance analysis we established a ...200010951583
positive- and negative-acting kruppel-like transcription factors bind a transforming growth factor beta control element required for expression of the smooth muscle cell differentiation marker sm22alpha in vivo.transforming growth factor beta (tgf-beta) is implicated in the regulation of smooth muscle cell (smc) differentiation. we previously identified a novel tgf-beta control element (tce) in the promoters of smc differentiation marker genes, including alpha-smooth muscle actin and sm22alpha. in this study, the importance of the tce in regulation of sm22alpha gene expression in vivo was investigated by mutating it within the context of a mouse sm22alpha promoter-lacz transgenic construct. mutation of ...200010954723
a role for a timp-3-sensitive, zn(2+)-dependent metalloprotease in mammalian gamete membrane fusion.during fertilization, sperm and egg plasma membranes adhere and then fuse by a mechanism that is not well understood. zinc metalloproteases are necessary for some intercellular fusion events, for instance, cell-cell fusion in yeast. in this study we tested the effects of class-specific and family-specific protease inhibitors on mouse gamete fusion. capacitated, acrosome-reacted sperm and zona-free eggs were used in assays designed to define the effects of inhibitors on sperm-egg plasma membrane ...200010964469
mouse prenylated rab acceptor is a novel golgi membrane protein.we have cloned a mouse prenylated rab acceptor (mpra), which interacts with various rab proteins in the yeast two-hybrid system. this study investigated its intracellular localization and characterized the localization signal. the mpra was found to be an integral membrane protein that was localized to the golgi complex at steady state as determined by confocal fluorescence microscopy. with green fluorescent protein attached to the n-terminus of mpra, the fusion protein was expressed in bhk cells ...200010964695
cbln3, a novel member of the precerebellin family that binds specifically to cbln1.precerebellin (cbln1) is the precursor of the brain-specific hexadecapeptide cerebellin. although cerebellin has properties of a conventional neuropeptide, its function is controversial because cbln1 has structural features characteristic of circulating atypical collagens. cbln1 is related to the three subunits of the complement c1q complex. therefore, we hypothesized that cbln1 participated in analogous heteromeric complexes with precerebellin-related proteins. using lexa-cbln1 as bait in a yea ...200010964938
characterization of tcl, a new gtpase of the rho family related to tc10 andccdc42.gtpases of the rho family control a wide variety of cellular processes such as cell morphology, motility, proliferation, differentiation, and apoptosis. we report here the characterization of a new rho member, which shares 85% and 78% amino acid similarity to tc10 and cdc42, respectively. this gtpase, termed as tc10-like (tcl) is encoded by an unexpectedly large locus, made of five exons spanning over 85 kilobases on human chromosome 14. tcl mrna is 2.5 kilobases long and is mainly expressed in ...200010967094
mdm2 interacts with the c-terminus of the catalytic subunit of dna polymerase epsilon.mdm2 is induced by p53 in response to cellular insults such as dna damage and can have effects upon the cell cycle that are independent or downstream of p53. we used a yeast two-hybrid screen to identify proteins that bind to mdm2 and which therefore might be involved in these effects. we found that mdm2 can bind to the c-terminus of the catalytic subunit of dna polymerase epsilon (dna pol epsilon), to a region that is known to be essential in yeast. in an in vitro system we confirmed that mdm2 ...200010982879
fatz, a filamin-, actinin-, and telethonin-binding protein of the z-disc of skeletal muscle.we report the identification and characterization of a novel 32-kda protein expressed in skeletal muscle and located in the z-disc of the sarcomere. we found that this protein binds to three other z-disc proteins; therefore, we have named it fatz, gamma-filamin/abp-l, alpha-actinin and telethonin binding protein of the z-disc. from yeast two-hybrid experiments we are able to show that the sr3-sr4 domains of alpha-actinin 2 are required to bind the cooh-terminal region of the fatz as does gamma-f ...200010984498
the werner syndrome. a model for the study of human aging.human aging is a complex process that leads to the gradual deterioration of body functions with time. various models to approach the study of aging have been launched over the years such as the genetic analysis of life span in the yeast s. cerevisiae, the worm c. elegans, the fruitfly, and mouse, among others. in human models, there have been extensive efforts using replicative senescence, the study of centenerians, comparisons of young versus old at the organismal, cellular, and molecular level ...200010911957
interaction of the tau2 transcriptional activation domain of glucocorticoid receptor with a novel steroid receptor coactivator, hic-5, which localizes to both focal adhesions and the nuclear matrix.hic-5 (hydrogen peroxide-inducible clone-5) is a focal adhesion protein that is involved in cellular senescence. in the present study, a yeast two-hybrid screen identified hic-5 as a protein that interacts with a region of the glucocorticoid receptor that includes a nuclear matrix-targeting signal and the tau2 transcriptional activation domain. in transiently transfected mammalian cells, overexpression of hic-5 potentiated the activation of reporter genes by all steroid receptors, excluding the ...200010848625
14-3-3 protein family members have a regulatory role in retinoic acid-mediated induction of cytokeratins in f9 cells.we have found that the expression of five 14-3-3 protein isoforms is induced during the retinoic acid (ra)-mediated differentiation of mouse embryonal carcinoma f9 cells. the induced expression of the 14-3-3 proteins is presumed to have a role in enhancing the mitogen-activated protein kinase (mapk) activity during ra-mediated f9 cell differentiation, because using genetically engineered budding yeast we showed that these isoforms enhanced the signaling in the mapk cascade mainly through the int ...200011010814
fine-scale comparative mapping of the human 7q11.23 region and the orthologous region on mouse chromosome 5g: the low-copy repeats that flank the williams-beuren syndrome deletion arose at breakpoint sites of an evolutionary inversion(s).williams-beuren syndrome (wbs) is a developmental disorder caused by haploinsufficiency for genes deleted in chromosome band 7q11.23. a common deletion including at least 16-17 genes has been defined in the great majority of patients. we have completed a physical and transcription map of the wbs region based on analysis of high-throughput genome sequence data and assembly of a bac/pac/yac contig, including the characterization of large blocks of gene-containing low-copy-number repeat elements th ...200011013070
the protein aly links pre-messenger-rna splicing to nuclear export in metazoans.in metazoans, most pre-messenger rnas contain introns that are removed by splicing. the spliced mrnas are then exported to the cytoplasm. recent studies showed that splicing promotes efficient mrna export, but the mechanism for coupling these two processes is not known. here we show that aly, the metazoan homologue of the yeast mrna export factor yralp (ref. 2), is recruited to messenger ribonucleoprotein (mrnp) complexes generated by splicing. in contrast, aly does not associate with mrnps asse ...200011014198
the apoptosis mediator mdap-3 is a novel member of a conserved family of mitochondrial proteins.programmed cell death is essential for organ development and regeneration. to identify molecules relevant for this process, full length cdna cloning of a short, developmentally regulated murine cdna fragment, merm-3, was performed and showed a 1.7 kb mrna encoding a 45 kda protein with an atp/gtp binding motive (p-loop). sequence analysis revealed an 82% amino acid identity to the human death associated protein 3 (hdap-3), a positive mediator of apoptosis. the full length sequence being the muri ...200011017876
[pharmacognosical study on secondary metabolites].clonal micropropagation on various medicinal plants was set up resulting in the regenerated plants which possessed a homogeneous quality. the ratio of hapten to bovine serum albumin (bsa) in an antigen conjugate was determined by matrix-assisted laser desorption/ionization of mass spectrometry. a hybridoma secreting monoclonal antibody (mab) was produced by fusing splenocytes immunized with an antigene-bsa conjugate with mouse myeloma cells. competitive enzyme-linked immunosorbent assay (elisa) ...200011019648
yeast two-hybrid system identifies the ubiquitin-conjugating enzyme mubc9 as a potential partner of mouse dac.using a yeast two hybrid system and pull-down assays we demonstrate that mouse dac (mdac) specifically binds to mouse ubiquitin-conjugating enzyme mubc9. in contrast to a direct interaction between drosophila dachshund (dac) and eyes absent (eya)gene products, we cannot detect by the same methods that mdac binds to meya2, a functional mouse homologue of the drosophila eya. immunostaining of various cell lines that were transfected with mdac reveals that mdac protein is found predominantly in the ...200011025202
a putative oncogenic role for mpp11 in head and neck squamous cell cancer.genetic alterations of chromosome 7 are common in human cancer. furthermore, previous studies have supported the presence of a gene important in a broad range of cancers at 7q22-31.1. there is evidence that supports an oncogenic function for this putative gene, as well as evidence that supports a tumor suppressive role. in this study, we used a cross-species candidate gene approach in combination with physical mapping to identify mpp11 as a candidate for the putative cancer-related activity at 7 ...200011034098
activating protein-1, nuclear factor-kappab, and serum response factor as novel target molecules of the cancer-amplified transcription coactivator asc-2.asc-2 was recently discovered as a cancer-amplified transcription coactivator molecule of nuclear receptors, which interacts with multifunctional transcription integrators steroid receptor coactivator-1 (src-1) and creb-binding protein (cbp)/p300. herein, we report the identification of three mitogenic transcription factors as novel target molecules of asc-2. first, the c-terminal transactivation domain of serum response factor (srf) was identified among a series of asc-2-interacting proteins fr ...200010847592
a family of lim-only transcriptional coactivators: tissue-specific expression and selective activation of creb and crem.transcription factors of the creb family control the expression of a large number of genes in response to various signaling pathways. regulation mediated by members of the creb family has been linked to various physiological functions. classically, activation by creb is known to occur upon phosphorylation at an essential regulatory site (ser133 in creb) and the subsequent interaction with the ubiquitous coactivator creb-binding protein (cbp). however, the mechanism by which selectivity is achiev ...200011046156
disruption of mouse polymerase zeta (rev3) leads to embryonic lethality and impairs blastocyst development in vitro.multiple dna polymerases exist in eukaryotes. polymerases alpha, delta and epsilon are mainly responsible for chromosomal dna replication in the nucleus and are required for proliferation. in contrast, the repair polymerases beta and eta are not essential for cellular proliferation in yeast or mice, but a lack of either polymerase can lead, respectively, to defects in base excision repair or the ability to replicate past lesions induced by ultraviolet (uv) radiation [1-3]. here, we have focused ...200011050391
overexpression of da41 in v-ha-ras-3y1 cells causes growth suppression.we have recently found that da41 exhibits marked homology with mouse plic-1, plic-2, frog xdrp1 and yeast dsk2. xdrp1 has been shown to be associated with cyclin a, and blocks cell division of frog embryo. in the present study, we examined the biological role(s) of da41 in mammalian cells by overexpressing it in v-ha-ras-transformed 3y1 cells (ras-3y1). transfectants which expressed a high level of da41 mrna exhibited a decrease in growth rate, a reduction in saturation density, and a suppressio ...200011050468
a sequence-ready pac contig of a 550-kb region on rat chromosome 4 including the diabetes susceptibility gene lyp.the lyp locus controls diabetes development in rats. the diabetogenic allele in diabetes-prone bb rats is responsible for t cell lymphopenia characterized by the absence of regulatory t cells. we present refined genetic and radiation hybrid maps of the lyp region on rat chromosome 4, a single 800-kb rat yeast artificial chromosome and a rat p1-derived artificial chromosome (pac) contig corresponding to approximately 550 kb, both encompassing the entire candidate region. the contig, consisting of ...200011056048
association of l-type calcium channels with a vacuolar h(+)-atpase g2 subunit.the class c l-type calcium (ca(2+)) channels have been implicated in many important physiological processes. here, we have identified a mouse vacuolar h(+)-atpase (v-atpase) g2 subunit protein that bound to the c-terminal domain of the pore-forming alpha(1c) subunit using a yeast two-hybrid screen. protein-protein interaction between the v-atpase g subunit and the alpha(1c) subunit was confirmed using in vitro gst pull-down assays and coimmunoprecipitation from intact cells. moreover, treatment ...200011062002
genome scan identifies a locus affecting gamma-globin level in human beta-cluster yac transgenic mice.genetic factors affecting postnatal gamma-globin expression--a major modifier of the severity of both beta-thalassemia and sickle cell anemia--have been difficult to study. this is especially so in mice, an organism lacking a globin gene with an expression pattern equivalent to that of human gamma-globin. to model the human beta-cluster in mice, with the goal of screening for loci affecting human gamma-globin expression in vivo, we introduced a human beta-globin cluster yac transgene into the ge ...200011063260
a fission yeast homolog of int-6, the mammalian oncoprotein and eif3 subunit, induces drug resistance when overexpressed.through a screen to identify genes that induce multi-drug resistance when overexpressed, we have identified a fission yeast homolog of int-6, a component of the human translation initiation factor eif3. disruption of the murine int-6 gene by mouse mammary tumor virus (mmtv) has been implicated previously in tumorigenesis, although the underlying mechanism is not yet understood. fission yeast int6 was shown to interact with other presumptive components of eif3 in vivo, and was present in size fra ...200011071922
diacylglycerol kinase zeta in hypothalamus interacts with long form leptin receptor. relation to dietary fat and body weight regulation.leptin and its long form receptor, ob-rb, in hypothalamic nuclei play a key role in regulating energy balance. the mutation of ob-rb into one of its natural variants, ob-ra, results in severe obesity in rodents. we demonstrate here that diacylglycerol kinase zeta (dgkzeta) interacts, via its ankyrin repeats, with the cytoplasmic portion of ob-rb in yeast two-hybrid systems, in protein precipitation experiments in vitro and in vivo. it does not interact, however, with the short form, ob-ra, which ...200111078732
vezatin, a novel transmembrane protein, bridges myosin viia to the cadherin-catenins complex.defects in myosin viia are responsible for deafness in the human and mouse. the role of this unconventional myosin in the sensory hair cells of the inner ear is not yet understood. here we show that the c-terminal ferm domain of myosin viia binds to a novel transmembrane protein, vezatin, which we identified by a yeast two-hybrid screen. vezatin is a ubiquitous protein of adherens cell-cell junctions, where it interacts with both myosin viia and the cadherin-catenins complex. its recruitment to ...200011080149
the murine dscr1-like (down syndrome candidate region 1) gene family: conserved synteny with the human orthologous genes.a recently recognized gene family, conserved from yeast to humans, includes down syndrome candidate region 1 gene (dscr1), adapt78 (recognized as the hamster ortholog of the dscr1 isoform 4), zaki-4 (renamed dscr1-like 1, dscr1l1) and dscr1l2 (a novel gene on human chromosome 1), along with yeast and c. elegans single members (strippoli p., lenzi l., petrini m., carinci p., zannotti m., 2000. a new gene family including dscr1 (down syndrome candidate region 1) and zaki-4: characterization from y ...200011080588
the human preprotachykinin-a gene promoter has been highly conserved and can drive human-like marker gene expression in the adult mouse cns.toward an understanding of the mechanisms controlling preprotachykinin-a (ppta) transcription, we introduced a 380-kb human yeast artificial chromosome containing the ppta gene tagged with the beta-galactosidase gene into transgenic mice. this resulted in a pattern of lacz expression in the central nervous system (cns) remarkably similar to that reported for ppta mrna in the rat. however, the human gene drove expression in areas of the mouse cns not associated with strong ppta expression in rode ...200011083923
arginine-rich peptides. an abundant source of membrane-permeable peptides having potential as carriers for intracellular protein delivery.a basic peptide derived from human immunodeficiency virus (hiv)-1 tat protein (positions 48-60) has been reported to have the ability to translocate through the cell membranes and accumulate in the nucleus, the characteristics of which are utilized for the delivery of exogenous proteins into cells. based on the fluorescence microscopic observations of mouse macrophage raw264.7 cells, we found that various arginine-rich peptides have a translocation activity very similar to tat-(48-60). these inc ...200111084031
the pea3 ets transcription factor comprises multiple domains that regulate transactivation and dna binding.pea3, a member of the ets family of transcription factors, is a nuclear phosphoprotein capable of activating transcription. mouse pea3 comprises 480 amino acids and bears an approximately 85-amino acid ets domain near its carboxyl terminus. whereas analyses of bacterially expressed pea3 revealed that the ets domain is required for sequence-specific dna binding, little is known of the functional domains in the protein required for its activity in mammalian cells. to this end, we defined the locat ...200111096072
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 ...200011102516
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 ...200011106249
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 ...200011106738
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