Publications

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conditional stimulation of type v and vi adenylyl cyclases by g protein betagamma subunits.in a yeast two-hybrid screen of mouse brain cdna library, using the n-terminal region of human type v adenylyl cyclase (hacv) as bait, we identified g protein beta2 subunit as an interacting partner. additional yeast two-hybrid assays showed that the gbeta(1) subunit also interacts with the n-terminal segments of hacv and human type vi adenylyl cyclase (hacvi). in vitro adenylyl cyclase (ac) activity assays using membranes of sf9 cells expressing hacv or hacvi showed that gbetagamma subunits enh ...200717110384
murine pif1 interacts with telomerase and is dispensable for telomere function in vivo.pif1 is a 5'-to-3' dna helicase critical to dna replication and telomere length maintenance in the budding yeast saccharomyces cerevisiae. scpif1 is a negative regulator of telomeric repeat synthesis by telomerase, and recombinant scpif1 promotes the dissociation of the telomerase rna template from telomeric dna in vitro. in order to dissect the role of mpif1 in mammals, we cloned and disrupted the mpif1 gene. in wild-type animals, mpif1 expression was detected only in embryonic and hematopoieti ...200717130244
the nucleolar protein surf-6 is essential for viability in mouse nih/3t3 cells.surf-6 is a bona fide nucleolar protein comprising an evolutionary conserved family that extends from human to yeast. the expression of the mammalian surf-6 has been recently found to be regulated during the cell cycle. in order to determine the importance of surf-6 in mammalian cells, we applied the tet-on system to regulate conditionally, in response to tetracycline, the expression of an antisense rna (asrna) that targets surf-6 mrna in mouse nih/3t3 cells. induced surf-6 asrna caused an effec ...200717086444
impact of transcriptional properties on essentiality and evolutionary rate.we characterized general transcriptional activity and variability of eukaryotic genes from global expression profiles of human, mouse, rat, fly, plants, and yeast. the variability shows a higher degree of divergence between distant species, implying that it is more closely related to phenotypic evolution, than the activity. more specifically, we show that transcriptional variability should be a true indicator of evolutionary rate. if we rule out the effect of translational selection, which seems ...200717057246
yeast sphingolipids do not need to contain very long chain fatty acids.synthesis of vlcfas (very long chain fatty acids) and biosynthesis of dhs (dihydrosphingosine) both are of vital importance for saccharomyces cerevisiae. the bulk of vlcfas and dhs are used for ceramide synthesis by the lag1p (longevity-assurance gene 1)/lac1p (longevity-assurance gene cognate 1)/lip1p (lag1p/lac1p interacting protein) ceramide synthase. lag1 and lac1 are redundant but lip1 is essential. here we show that 4delta (lag1deltalac1deltaypc1deltaydc1delta) cells devoid of all known en ...200716987101
inactive full-length p53 mutants lacking dominant wild-type p53 inhibition highlight loss of heterozygosity as an important aspect of p53 status in human cancers.over 1000 different mutants of the tumor suppressor protein p53 with one amino acid change in the core domain have been reported in human cancers. in mouse knock-in models, two frequent mutants displayed loss of wild-type (wt) p53 function, inhibition of wt p53 and wt p53-independent gain of function. the remaining mutants have been systematically characterized for loss of wt p53 function, but not other phenotypes. we report the concomitant assessment of loss of function and interference with wt ...200716861262
cloning, analysis and functional annotation of expressed sequence tags from the earthworm eisenia fetida.eisenia fetida, commonly known as red wiggler or compost worm, belongs to the lumbricidae family of the annelida phylum. little is known about its genome sequence although it has been extensively used as a test organism in terrestrial ecotoxicology. in order to understand its gene expression response to environmental contaminants, we cloned 4032 cdnas or expressed sequence tags (ests) from two e. fetida libraries enriched with genes responsive to ten ordnance related compounds using suppressive ...200718047730
overview of gene targeting by homologous recombination.the analysis of mutant organisms and cell lines is important in determining the function of specific proteins. recent technological advances in gene targeting by homologous recombination in mammalian systems enable the production of mutants in any desired gene, and can be used to produce mutant mouse strains and mutant cell lines. the yeast flp/frt recombinase system and bacteriophage recombinases such as cre and its recognition sequence, loxp, allow spatial and temporal control of knockouts. th ...200718428657
accentuate the negative: proteome comparisons using the negative proteome database.the availability of complete genome sequence information for diverse organisms including model genetic organisms has ushered in a new era of protein sequence comparisons making it possible to search for commonalities among entire proteomes using the basic local alignment search tool (blast). although the identification and analysis of proteins shared by humans and model organisms has proven an invaluable tool to understanding gene function, the sets of proteins unique to a given model organism's ...200718820470
munc18a scaffolds snare assembly to promote membrane fusion.munc18a is an sm protein required for snare-mediated fusion. the molecular details of how munc18a acts to enhance neurosecretion have remained elusive. here, we use in vitro fusion assays to characterize how specific interactions between munc18a and the neuronal snares enhance the rate and extent of fusion. we show that munc18a interacts directly and functionally with the preassembled t-snare complex. analysis of munc18a point mutations indicates that munc18a interacts with helix c of the syntax ...200818829865
the scx/imac enrichment approach for global phosphorylation analysis by mass spectrometry.the success in profiling the phosphoproteome by mass spectrometry-based proteomics has been intimately related to the availability of methods that selectively enrich for phosphopeptides. to this end, we describe a protocol that combines two sequential enrichment steps. first, strong cation exchange (scx) chromatography separates peptides by solution charge. phosphate groups contribute to solution charge by adding a negative charge at ph 2.7. therefore, at that ph, phosphopeptides are expected to ...200818833199
impaired dna damage response, genome instability, and tumorigenesis in sirt1 mutant mice.in lower eukaryotes, sir2 serves as a histone deacetylase and is implicated in chromatin silencing, longevity, and genome stability. here we mutated the sirt1 gene, a homolog of yeast sir2, in mice to study its function. we show that a majority of sirt1 null embryos die between e9.5 and e14.5, displaying altered histone modification, impaired dna damage response, and reduced ability to repair dna damage. we demonstrate that sirt1(+/-);p53(+/-) mice develop tumors in multiple tissues, whereas act ...200818835033
'knock, and it shall be opened': knocking out and knocking in to reveal mechanisms of disease and novel therapies.recent significant advances in molecular biology have generated genetically modified bacteria, yeast, nematodes, fruit flies, and fish. however, it is the genetic modification of mammalian model organisms, particularly the mouse, that has the greatest potential to shed light on human development, physiology and pathology in ways that have significant implications for neonatal and paediatric clinical practice. here, we review some of the techniques for knocking out (inactivating), mutating and kn ...200818838236
sirt1 gain of function increases energy efficiency and prevents diabetes in mice.in yeast, worms, and flies, an extra copy of the gene encoding the sirtuin sir2 increases metabolic efficiency, as does administration of polyphenols like resveratrol, thought to act through sirtuins. but evidence that sirtuin gain of function results in increased metabolic efficiency in mammals is limited. we generated transgenic mice with moderate overexpression of sirt1, designed to mimic the sirtuin gain of function that improves metabolism in c. elegans. these mice exhibit normal insulin se ...200818840364
engineering higher affinity t cell receptors using a t cell display system.the t cell receptor (tcr) determines the cellular response to antigens, which are presented on the surface of target cells in the form of a peptide bound to a product of the major histocompatibility complex (pepmhc). the response of the t cell depends on the affinity of the tcr for the pepmhc, yet many tcrs have been shown to be of low affinity, and some naturally occurring t cell responses are poor due to low affinities. accordingly, engineering the tcr for increased affinity for pepmhc, partic ...200818854190
mink is a rap2 effector for phosphorylation of the postsynaptic scaffold protein tanc1.rap1 and rap2 are similar ras-like g proteins but perform distinct functions. by the affinity chromatography/mass-spectrometry approach and the yeast two-hybrid screening, we identified misshapen/niks-related kinase (mink) as a novel rap2-interacting protein that does not interact with rap1 or ras. mink is a member of the ste20 group of mitogen-activated protein kinase kinase kinase kinases. the interaction between mink and rap2 was gtp-dependent and required phe39 within the effector region of ...200818930710
bacteriophage mu integration in yeast and mammalian genomes.genomic parasites have evolved distinctive lifestyles to optimize replication in the context of the genomes they inhabit. here, we introduced new dna into eukaryotic cells using bacteriophage mu dna transposition complexes, termed 'transpososomes'. following electroporation of transpososomes and selection for marker gene expression, efficient integration was verified in yeast, mouse and human genomes. although mu has evolved in prokaryotes, strong biases were seen in the target site distribution ...200818953026
ubc9-mediated sumoylation leads to transcriptional repression of irf-1.to characterize the regulatory mechanism of the interferon regulatory factor (irf) family, we performed yeast two-hybrid screening with irf-2 and isolated the small ubiquitin-related modifier (sumo)-conjugating enzyme ubc9, which also interacts with other irf family members irf-1 and icsbp. subsequent assays indicated that among the irf family members, only irf-1 interacts with sumo-1 through its transcriptional activation domain. the interaction between irf-1 and sumo-1 was confirmed in vitro b ...200818955028
sequence space coverage, entropy of genomes and the potential to detect non-human dna in human samples.genomes store information for building and maintaining organisms. complete sequencing of many genomes provides the opportunity to study and compare global information properties of those genomes.200818973670
predicting the proportion of essential genes in mouse duplicates based on biased mouse knockout genes.in the yeast or nematode, the proportion of essential genes in duplicates is lower than in singletons (single-copy genes), due to the functional redundancy. one may expect that it should be the same in the mouse genome. however, based on the publicly available mouse knockout data, it was observed that the proportion of essential genes in duplicates is similar to that in singletons. the most straightforward interpretation, as claimed in a recent study, is that duplicate genes may have a negligibl ...200819005716
restoration of electron transport without proton pumping in mammalian mitochondria.we have restored the coq oxidative capacity of mouse mtdna-less cells (rho degrees cells) by transforming them with the alternative oxidase aox of emericella nidulans. cotransforming rho degrees cells with the nadh dehydrogenase of saccharomyces cerevisiae, ndi1 and aox recovered the nadh dh/coq reductase and the coq oxidase activities. coq oxidation by aox reduces the dependence of rho degrees cells on pyruvate and uridine. coexpression of aox and ndi1 further improves the recycling of nad(+). ...200819020091
pcna interacts with prox1 and represses its transcriptional activity.prox1 is a transcription factor which can function either as a transcriptional activator, transcriptional repressor or a transcriptional corepressor. this paper seeks to better understand the role of protein-protein interactions in this multitude of functions.200819023449
vac14 nucleates a protein complex essential for the acute interconversion of pi3p and pi(3,5)p(2) in yeast and mouse.the signalling lipid pi(3,5)p(2) is generated on endosomes and regulates retrograde traffic to the trans-golgi network. physiological signals regulate rapid, transient changes in pi(3,5)p(2) levels. mutations that lower pi(3,5)p(2) cause neurodegeneration in human patients and mice. the function of vac14 in the regulation of pi(3,5)p(2) was uncharacterized previously. here, we predict that yeast and mammalian vac14 are composed entirely of heat repeats and demonstrate that vac14 exerts an effect ...200819037259
insig regulates hmg-coa reductase by controlling enzyme phosphorylation in fission yeast.insig functions as a central regulator of cellular cholesterol homeostasis by controlling activity of hmg-coa reductase (hmgr) in cholesterol synthesis. insig both accelerates the degradation of hmgr and suppresses hmgr transcription through the srebp-scap pathway. the fission yeast schizosaccharomyces pombe encodes homologs of insig, hmgr, srebp, and scap, called ins1(+), hmg1(+), sre1(+), and scp1(+). here, we characterize fission yeast insig and demonstrate that ins1 is dedicated to regulatio ...200819041767
diffusion magnetic resonance imaging: an imaging treatment response biomarker to chemoradiotherapy in a mouse model of squamous cell cancer of the head and neck.for the treatment of squamous cell cancer of the head and neck (scchn), the assessment of treatment response is traditionally accomplished by volumetric measurements and has been suggested to be prognostic for an eventual response to treatment. an early evaluation response during the course of radiation therapy could provide an opportunity to tailor treatment to individual patients. diffusion magnetic resonance imaging (mri) allows for the quantification of tissue water diffusion values, thus tr ...200819043529
recruitment of rna polymerase ii cofactor pc4 to dna damage sites.the multifunctional nuclear protein positive cofactor 4 (pc4) is involved in various cellular processes including transcription, replication, and chromatin organization. recently, pc4 has been identified as a suppressor of oxidative mutagenesis in escherichia coli and saccharomyces cerevisiae. to investigate a potential role of pc4 in mammalian dna repair, we used a combination of live cell microscopy, microirradiation, and fluorescence recovery after photobleaching analysis. we found a clear ac ...200819047459
regulation of forkhead transcription factor foxo3a contributes to calorie restriction-induced prevention of alzheimer's disease-type amyloid neuropathology and spatial memory deterioration.forkhead transcription factor foxo3a, also known as daf-16 in caenorhabditis elegans, is a key regulator of the insulin receptor (ir)/insulin-like growth factor-i signaling pathway mediated extension of life span in worms and yeast. in this study, we report that calorie restriction (cr)-mediated activation of the ir signaling pathway leads to hyperphosphorylation of foxo3a transcription factor and, consequently, its exclusion from the nucleus. this inactivation of foxo3a activity is correlated w ...200819076455
buffering by gene duplicates: an analysis of molecular correlates and evolutionary conservation.one mechanism to account for robustness against gene knockouts or knockdowns is through buffering by gene duplicates, but the extent and general correlates of this process in organisms is still a matter of debate. to reveal general trends of this process, we provide a comprehensive comparison of gene essentiality, duplication and buffering by duplicates across seven bacteria (mycoplasma genitalium, bacillus subtilis, helicobacter pylori, haemophilus influenzae, mycobacterium tuberculosis, pseudo ...200819087332
research on animal model organisms funded by the european commission's framework programmes.recognising the crucial role of model organisms in exploring the causes of human disease and in developing safe treatments, the european commission has invested euro180 million in collaborative research projects on model organisms since 2002. further financial support is planned for the future. projects supported by the european commission are playing an important role in structuring the research landscape in europe and creating the knowledge base to understand health and disease. furthermore, t ...200819093026
sirt7-dependent inhibition of cell growth and proliferation might be instrumental to mediate tissue integrity during aging.mammalian sirtuins, sirt1-sirt7, are recently discovered regulatory proteins, which play decisive roles in cellular metabolism, stress resistance, and proliferation. sirtuins are homologs of the founder member of the sirtuin family, the yeast sir2. sir2 encodes a nad(+)-dependent histone deacetylase and its overexpression extends the lifespan through silencing of specific chromatin regions. lifespan extension by sir2 homologs was also demonstrated in more complex species such as c. elegans and d ...200819261981
antihypertensive drug guanabenz is active in vivo against both yeast and mammalian prions.prion-based diseases are incurable transmissible neurodegenerative disorders affecting animals and humans.200818431471
conserved co-expression for candidate disease gene prioritization.genes that are co-expressed tend to be involved in the same biological process. however, co-expression is not a very reliable predictor of functional links between genes. the evolutionary conservation of co-expression between species can be used to predict protein function more reliably than co-expression in a single species. here we examine whether co-expression across multiple species is also a better prioritizer of disease genes than is co-expression between human genes alone.200818433471
regulation of apoptosis and neurite extension by fkbp38 is required for neural tube formation in the mouse.fkbp38 (also known as fkbp8) is a transmembrane chaperone protein that inhibits apoptosis by recruiting the anti-apoptotic proteins bcl-2 and bcl-x(l) to mitochondria. we have now generated mice harboring a loss-of-function mutation in fkbp38. the fkbp38(-/-) mice die soon after birth manifesting defects in neural tube closure in the thoraco-lumbar-sacral region (spina bifida) as well as skeletal defects including scoliosis, rib deformities, club foot and curled tail. the neuroepithelium is diso ...200818459960
live yeast cell derivative induces c-fos expression in thp-1 monocytes.live yeast cell derivative is a medicinal extract of saccharomyces cerevisiae that has demonstrated efficacy in improving the rate and quality of wound healing in mouse and human systems. however, the mechanisms by which lycd promotes healing are largely uncharacterized. in this report, we demonstrate that lycd has effects on the transcriptional profile of the human monocytic cell line thp-1. thirty minute exposures of thp-1 cells with lycd induced a 6 to 44-fold, dose-dependent increase in the ...200818473919
advanced method for high-throughput expression of mutated eukaryotic membrane proteins in saccharomyces cerevisiae.crystallization of eukaryotic membrane proteins is a challenging, iterative process. the protein of interest is often modified in an attempt to improve crystallization and diffraction results. to accelerate this process, we took advantage of a gfp-fusion yeast expression system that uses pcr to direct homologous recombination and gene cloning. we explored the possibility of employing more than one pcr fragment to introduce various mutations in a single step, and found that when up to five pcr fr ...200818474222
amino acid biases in the n- and c-termini of proteins are evolutionarily conserved and are conserved between functionally related proteins.analysis of the arabidopsis thaliana, saccharomyces cerevisiae, mus musculus, escherichia coli, bacillus subtilis, thermoplasma acidophilum, and sulfolobus tokodaii genomes demonstrate that many amino acid biases occur at the n- and c-termini of proteins, a statistically significant number of these biases are evolutionarily conserved, and these biases occur in amino acids beyond the first and last five amino acids. analyses designed to shed light on the mechanism causing amino acid biases sugges ...200818478318
lkb1 and lung cancer: more than the usual suspects.often, the problem in cancer research is figuring out how a gene or pathway works in regulating cellular transformation. the question of what ras activates or pten inhibits have been classic dilemmas of modern cancer biology. in these cases, biochemical and genetic studies have provided us with a fairly clear picture of the cancer relevant functions of these genes. for lkb1, a more recently identified human tumor suppressor gene, however, the problem is different. this serine-threonine kinase th ...200818483235
the macrophage-inducible c-type lectin, mincle, is an essential component of the innate immune response to candida albicans.the recognition of carbohydrate moieties by cells of the innate immune system is emerging as an essential element in antifungal immunity, but despite the number and diversity of lectins expressed by innate immune cells, few carbohydrate receptors have been characterized. mincle, a c-type lectin, is expressed predominantly on macrophages, and is here shown to play a role in macrophage responses to the yeast candida albicans. after exposure to the yeast in vitro, mincle localized to the phagocytic ...200818490740
hubba: hub objects analyzer--a framework of interactome hubs identification for network biology.one major task in the post-genome era is to reconstruct proteomic and genomic interacting networks using high-throughput experiment data. to identify essential nodes/hubs in these interactomes is a way to decipher the critical keys inside biochemical pathways or complex networks. these essential nodes/hubs may serve as potential drug-targets for developing novel therapy of human diseases, such as cancer or infectious disease caused by emerging pathogens. hub objects analyzer (hubba) is a web-bas ...200818503085
ulk1 plays a critical role in the autophagic clearance of mitochondria and ribosomes during reticulocyte maturation.production of a red blood cell's hemoglobin depends on mitochondrial heme synthesis. however, mature red blood cells are devoid of mitochondria and rely on glycolysis for atp production. the molecular basis for the selective elimination of mitochondria from mature red blood cells remains controversial. recent evidence suggests that clearance of both mitochondria and ribosomes, which occurs in reticulocytes following nuclear extrusion, depends on autophagy. here, we demonstrate that ulk1, a serin ...200818539900
construction of cell surface-engineered yeasts displaying antigen to detect antibodies by immunofluorescence and yeast-elisa.in order to detect monoclonal antibodies (mabs) from insufficient and unavailable human proteins, yeast cells were engineered to display human antigens on their surface and consequently endowed with the ability to specifically bind antibodies. thus, a fusion gene for the expression of the human proteasome subunit alpha 6 (hpsa6) and human profilin i (hproi) were assembled, respectively, with a his.tag marker at the c-terminal and displayed on yeast surface. with anti-his.tag mab as the primary a ...200818542951
gal4/uas transgenic tools and their application to zebrafish.the ability to regulate gene expression in a cell-specific and temporally restricted manner provides a powerful means to test gene function, bypass the action of lethal genes, label subsets of cells for developmental studies, monitor subcellular structures, and target tissues for selective ablation or physiological analyses. the galactose-inducible system of yeast, mediated by the transcriptional activator gal4 and its consensus uas binding site, has proven to be a highly successful and versatil ...200818554173
regulation of rna polymerase iii transcription by maf1 protein.maf1 was the first protein discovered to regulate polymerase iii rna in yeast and because it is evolutionarily conserved, a maf1 ortholog also serves to restrain transcription in mouse and human cells. understanding the mechanism of the regulation has been made possible by recent studies showing that maf1 is a nuclear/cytoplasmic protein whose subcellular distribution and hence negative regulation of pol iii transcription is mediated by the nutrient-sensing signaling pathways, tor and ras. under ...200818560610
evolution of closely linked gene pairs in vertebrate genomes.the orientation of closely linked genes in mammalian genomes is not random: there are more head-to-head (h2h) gene pairs than expected. to understand the origin of this enrichment in h2h gene pairs, we have analyzed the phylogenetic distribution of gene pairs separated by less than 600 bp of intergenic dna (gene duos). we show here that a lack of head-to-tail (h2t) gene duos is an even more distinctive characteristic of mammalian genomes, with the platypus genome as the only exception. in nonmam ...200818566020
ccr2 mediates conventional dendritic cell recruitment and the formation of bronchovascular mononuclear cell infiltrates in the lungs of mice infected with cryptococcus neoformans.pulmonary clearance of the encapsulated yeast cryptococcus neoformans requires the development of t1-type immunity. ccr2-deficient mice infected with c. neoformans develop a non-protective t2 immune response and persistent infection. the mechanisms responsible for this aberrant response are unknown. the objective of this study was to define the number, phenotype, and microanatomic location of dendritic cells (dc) residing within the lung of ccr2+/+ or ccr2-/- mice throughout a time course follow ...200818566428
the mammalian target of rapamycin complex 2 controls folding and stability of akt and protein kinase c.the target of rapamycin (tor), as part of the rapamycin-sensitive tor complex 1 (torc1), regulates various aspects of protein synthesis. whether tor functions in this process as part of torc2 remains to be elucidated. here, we demonstrate that mtor, sin1 and rictor, components of mammalian (m)torc2, are required for phosphorylation of akt and conventional protein kinase c (pkc) at the turn motif (tm) site. this torc2 function is growth factor independent and conserved from yeast to mammals. tm s ...200818566586
dumpy-30 family members as determinants of male fertility and interaction partners of metal-responsive transcription factor 1 (mtf-1) in drosophila.metal-responsive transcription factor 1 (mtf-1), which binds to metal response elements (mres), plays a central role in transition metal detoxification and homeostasis. a drosophila interactome analysis revealed two candidate dmtf-1 interactors, both of which are related to the small regulatory protein dumpy-30 (dpy-30) of the worm c. elegans. dpy-30 is the founding member of a protein family involved in chromatin modifications, notably histone methylation. mutants affect mating type in yeast an ...200818588663
an en masse phenotype and function prediction system for mus musculus.individual researchers are struggling to keep up with the accelerating emergence of high-throughput biological data, and to extract information that relates to their specific questions. integration of accumulated evidence should permit researchers to form fewer - and more accurate - hypotheses for further study through experimentation.200818613952
fermented guava leaf extract inhibits lps-induced cox-2 and inos expression in mouse macrophage cells by inhibition of transcription factor nf-kappab.the goal of this study was to elucidate the antiinflammatory activities of psidium guajava l. (guava) leaf. to improve the functionality of guava leaf, it was fermented with phellinus linteus mycelia, lactobacillus plantarum and saccharomyces cerevisiae. the ethanol extract from fermented guava leaf inhibited lipopolysaccharide (lps)-induced nitric oxide (no) and prostaglandin e(2) (pge(2)) production. western blot analysis showed that fermented guava leaf extract decreased lps-induced inducible ...200818618521
cloning and characterization of e-dlg, a novel splice variant of mouse homologue of the drosophila discs large tumor suppressor binds preferentially to sap102.membrane-associated guanylate kinases (maguks) act as scaffolds to coordinate signaling events through their multiple domains at the plasma membrane. the maguk sh3 domain is noncanonical and its function remains unclear. to identify potential binding partners of maguk sh3, the synapse-associated protein 102 (sap102) sh3 domain was used as bait in a yeast two-hybrid screen of a mouse embryonic cdna library. a mouse homologue of the drosophila discs large tumor suppressor (dlg, also known as sap97 ...200818618587
tripartite motif protein 32 facilitates cell growth and migration via degradation of abl-interactor 2.tripartite motif protein 32 (trim32) mrna has been reported to be highly expressed in human head and neck squamous cell carcinoma, but the involvement of trim32 in carcinogenesis has not been fully elucidated. in this study, we found by using yeast two-hybrid screening that trim32 binds to abl-interactor 2 (abi2), which is known as a tumor suppressor and a cell migration inhibitor, and we showed that trim32 mediates the ubiquitination of abi2. overexpression of trim32 promoted degradation of abi ...200818632609
jouberin localizes to collecting ducts and interacts with nephrocystin-1.joubert syndrome and related disorders are autosomal recessive multisystem diseases characterized by cerebellar vermis aplasia/hypoplasia, retinal degeneration and cystic kidney disease. there are five known genes; mutations of which give rise to a spectrum of renal cystic diseases the most common of which is nephronophthisis, a disorder characterized by early loss of urinary concentrating ability, renal fibrosis, corticomedullary cyst formation and renal failure. many of the proteins encoded by ...200818633336
expression and function of tetran, a new type of membrane transporter.in contrast to transport across basolateral membranes, the mechanism governing transport of organic anions across the luminal membranes of proximal tubules has remained unclear. we recently found tetracycline transporter-like protein (tetran), a human ortholog of yeast tpo1p that can transport anionic non-steroidal anti-inflammatory drugs (nsaids). in this study, we examine the expression and function of tetran. tetran mrna is expressed in various human tissues, including kidney. when overexpres ...200818638446
the response to a specific germinant by bacillus anthracis spores in primary mouse macrophages is modulated by a protein encoded on the pxo1 plasmid.a bacillus anthracis sterne pxo1 plasmid-encoded protein designated cot43 was found in coat extracts of purified spores. cot43 is a tetratricopeptide repeat domain protein related to those which function as phosphatases in the sporulation phosphorelay and as regulators of competence and pathogenic factors. the synthesis of cot43 began in the late exponential phase downstream from a sigmaa promoter (as mapped by race) and it was present at least until the formation of phase white endospores. ther ...200818654763
mistranslation-induced protein misfolding as a dominant constraint on coding-sequence evolution.strikingly consistent correlations between rates of coding-sequence evolution and gene expression levels are apparent across taxa, but the biological causes behind the selective pressures on coding-sequence evolution remain controversial. here, we demonstrate conserved patterns of simple covariation between sequence evolution, codon usage, and mrna level in e. coli, yeast, worm, fly, mouse, and human that suggest that all observed trends stem largely from a unified underlying selective pressure. ...200818662548
a novel tlr4-binding peptide that inhibits lps-induced activation of nf-kappab and in vivo toxicity.lipopolysaccharide (lps) is a component of the outer membrane of gram-negative bacteria. it is a ligand for toll-like receptor 4 (tlr4), which plays an essential role in innate immunity. macrophages and dendritic cells exposed to lps overproduce proinflammatory mediators, leading to septic shock. in this study, we screened for peptides that associate with tlr4 with a yeast two-hybrid screen using the human tlr4 extracellular domain as bait. a peptide (stm28) isolated from the screen inhibited lp ...200818700140
interaction assays in yeast and cultured cells confirm known and identify novel partners of the synaptic vesicle protein synaptophysin.synaptophysin (syp) is a major protein of neurotransmitter-containing vesicles spanning the membrane four times and contributing to various aspects of the synaptic vesicle cycle. the split-ubiquitin yeast two-hybrid system was used to characterize molecular interactions of membrane-bound, full-length murine syp. in this way, the known homophilic syp-syp association could be confirmed and heterophilic binding of syp to other tetraspan vesicle membrane proteins of the secretory carrier-associated ...200818706977
the putative alpha-1,2-mannosyltransferase afmnt1 of the opportunistic fungal pathogen aspergillus fumigatus is required for cell wall stability and full virulence.proteins entering the eukaryotic secretory pathway commonly are glycosylated. important steps in this posttranslational modification are carried out by mannosyltransferases. in this study, we investigated the putative alpha-1,2-mannosyltransferase afmnt1 of the human pathogenic mold aspergillus fumigatus. afmnt1 belongs to a family of enzymes that comprises nine members in saccharomyces cerevisiae but only three in a. fumigatus. a deltaafmnt1 mutant is viable and grows normally at 37 degrees c, ...200818708564
global alignment of multiple protein interaction networks with application to functional orthology detection.protein-protein interactions (ppis) and their networks play a central role in all biological processes. akin to the complete sequencing of genomes and their comparative analysis, complete descriptions of interactomes and their comparative analysis is fundamental to a deeper understanding of biological processes. a first step in such an analysis is to align two or more ppi networks. here, we introduce an algorithm, isorank, for global alignment of multiple ppi networks. the guiding intuition here ...200818725631
effect of starvation on transcriptomes of brain and liver in adult female zebrafish (danio rerio).we used microarray and quantitative real-time pcr (qrt-pcr) analyses in adult female zebrafish (danio rerio) to identify metabolic pathways regulated by starvation in the liver and brain. the transcriptome of whole zebrafish brain showed little response to 21 days of starvation. only agouti-related protein 1 (agrp1) significantly responded, with increased expression in brains of starved fish. in contrast, a 21-day period of starvation significantly downregulated 466 and upregulated 108 transcrip ...200818728227
diphthamide modification of eef2 requires a j-domain protein and is essential for normal development.the intracellular target of diphtheria toxin is a modified histidine residue, diphthamide, in the translation elongation factor, eef2 (also known as eft1). this enigmatic modification occurs in all eukaryotes and is produced in yeast by the action of five gene products, dph1 to dph5. sequence homologues of these genes are present in all sequenced eukaryotic genomes and, in higher eukaryotes, there is functional evidence for dph1, dph2, dph3 and dph5 acting in diphthamide biosynthesis. we identif ...200818765564
phosphorylation sites on tyr28 and the c-terminus of rad9 are required for inhibition of premature chromosomal condensation across the entire s phase.rad9 is required for the activity of the genotoxin-induced checkpoint signaling pathway to control cell cycle progress and maintain genomic stability. in the fission yeast s. pombe and chicken cells, the rad9 gene is essential for prevention of premature chromosomal condensation (pcc:s/m checkpoint control). however, precise features in the s/m checkpoint controlled by rad9 in mammalian cells are still not clear. in this study, mouse embryonic stem (es) cells with a targeted deletion of mrad9, t ...200818769057
mitop2: an integrative tool for the analysis of the mitochondrial proteome.mitochondria are crucial for normal cell metabolism and maintenance. mitochondrial dysfunction has been implicated in a spectrum of human diseases, ranging from rare monogenic to common multifactorial disorders. important for the understanding of organelle function is the assignment of its constituents, and although over 1,500 proteins are predicted to be involved in mammalian mitochondrial function, so far only about 900 are assigned to mitochondria with reasonable certainty. continuing efforts ...200818780189
sequential interval motif search: unrestricted database surveys of global ms/ms data sets for detection of putative post-translational modifications.tandem mass spectrometry is the prevailing approach for large-scale peptide sequencing in high-throughput proteomic profiling studies. effective database search engines have been developed to identify peptide sequences from ms/ms fragmentation spectra. since proteins are polymorphic and subject to post-translational modifications (ptm), however, computational methods for detecting unanticipated variants are also needed to achieve true proteome-wide coverage. different from existing "unrestrictiv ...200818788753
the role of calorie restriction and sirt1 in prion-mediated neurodegeneration.a central focus of aging research is to determine how calorie restriction (cr) extends lifespan and delays diseases of aging. sirt1, the mammalian ortholog of sir2 in yeast, is a longevity factor which mediates dietary restriction in diverse species. in addition, sirt1 plays a protective role in several models of neurodegenerative disease. we tested the role of sirt1 in mediating the effects of cr in a mouse model of prion disease. prion diseases are protein misfolding disorders of the central n ...200818799131
function and regulation of protein neddylation. 'protein modifications: beyond the usual suspects' review series.neddylation is the post-translational protein modification that is most closely related to ubiquitination. however, ubiquitination is known to regulate a myriad of processes in eukaryotic cells, whereas only a limited number of neddylation substrates have been described to date. here, we review the principles of protein neddylation and highlight the mechanisms that ensure the specificity of neddylation over ubiquitination. as numerous neddylation substrates probably remain to be discovered, we p ...200818802447
expression cloning of tmem16a as a calcium-activated chloride channel subunit.calcium-activated chloride channels (caccs) are major regulators of sensory transduction, epithelial secretion, and smooth muscle contraction. other crucial roles of caccs include action potential generation in characean algae and prevention of polyspermia in frog egg membrane. none of the known molecular candidates share properties characteristic of most caccs in native cells. using axolotl oocytes as an expression system, we have identified tmem16a as the xenopus oocyte cacc. the tmem16 family ...200818805094
a genomewide functional network for the laboratory mouse.establishing a functional network is invaluable to our understanding of gene function, pathways, and systems-level properties of an organism and can be a powerful resource in directing targeted experiments. in this study, we present a functional network for the laboratory mouse based on a bayesian integration of diverse genetic and functional genomic data. the resulting network includes probabilistic functional linkages among 20,581 protein-coding genes. we show that this network can accurately ...200818818725
localization of mouse mitochondrial sirt proteins: shift of sirt3 to nucleus by co-expression with sirt5.yeast silent information regulator 2 (sir2) is involved in extension of yeast longevity by calorie restriction, and sirt3, sirt4, and sirt5 are mammalian homologs of sir2 localized in mitochondria. we have investigated the localization of these three sirt proteins of mouse. sirt3, sirt4, and sirt5 proteins were localized in different compartments of the mitochondria. when sirt3 and sirt5 were co-expressed in the cell, localization of sirt3 protein changed from mitochondria to nucleus. these resu ...200818054327
unified mode of centromeric protection by shugoshin in mammalian oocytes and somatic cells.reductional chromosome segregation in germ cells, where sister chromatids are pulled to the same pole, accompanies the protection of cohesin at centromeres from separase cleavage. here, we show that mammalian shugoshin sgo2 is expressed in germ cells and is solely responsible for the centromeric localization of pp2a and the protection of cohesin rec8 in oocytes, proving conservation of the mechanism from yeast to mammals. however, this role of sgo2 contrasts with its mitotic role in protecting c ...200818084284
an intact unfolded protein response in trpt1 knockout mice reveals phylogenic divergence in pathways for rna ligation.unconventional mrna splicing by an endoplasmic reticulum stress-inducible endoribonuclease, ire1, is conserved in all known eukaryotes. it controls the expression of a transcription factor, hac1p/xbp-1, that regulates gene expression in the unfolded protein response. in yeast, the rna fragments generated by ire1p are ligated by trna ligase (trl1p) in a process that leaves a 2'-po4(2-) at the splice junction, which is subsequently removed by an essential 2'-phosphotransferase, tpt1p. however, ani ...200818094117
involvement of s100a6 (calcyclin) and its binding partners in intracellular signaling pathways. 200818155169
the cytoplasmic domain of chondrolectin interacts with the beta-subunit of rab geranylgeranyl transferase.mouse chondrolectin (chodl) was isolated out of the tail tip of four-day old 129/svj mice as a by-product of a pcr-based subtractive cdna library screening. the gene is predominantly expressed in adult skeletal muscle, heart, testes and lungs and in embryonic stadia. chodl is the mouse homologue of human chondrolectin (chodl), a gene that encodes for a type ia transmembrane protein and that is expressed in human testis, prostate, heart and skeletal muscle tissue. chodl-splice variants (chodl ( f ...200818161010
the n-end rule pathway is a sensor of heme.the conjugation of arginine, by arginyl-transferase, to n-terminal aspartate, glutamate or oxidized cysteine is a part of the n-end rule pathway of protein degradation. we report that arginyl-transferase of either the mouse or the yeast saccharomyces cerevisiae is inhibited by hemin (fe(3+)-heme). furthermore, we show that hemin inhibits arginyl-transferase through a redox mechanism that involves the formation of disulfide between the enzyme's cys-71 and cys-72 residues. remarkably, hemin also i ...200818162538
general transcription factor iia-gamma increases osteoblast-specific osteocalcin gene expression via activating transcription factor 4 and runt-related transcription factor 2.atf4 (activating transcription factor 4) is an osteoblast-enriched transcription factor that regulates terminal osteoblast differentiation and bone formation. atf4 knock-out mice have reduced bone mass (severe osteoporosis) throughout life. runx2 (runt-related transcription factor 2) is a runt domain-containing transcription factor that is essential for bone formation during embryogenesis and postnatal life. in this study, we identified general transcription factor iia gamma (tfiia gamma) as a r ...200818171674
the cloning of dmc1 cdnas and a comparative study of its expression in different ploidy cyprinid fishes.dmc1 (disrupted meiotic cdna) is a functionally specific gene, which was firstly discovered in yeast and then found to encode a protein required for homologous chromosome synapsis during the process of meiosis. in this investigation, we cloned the partial cdnas of dmc1 of diploid red crucian carp, japanese crucian carp, common carp, triploid crucian carp and allotetraploid hybrids by using a pair of degenerate primers based on the conservative sequence of amino acids of the dmc1 protein in yeast ...200818176789
a role for lengsin, a recruited enzyme, in terminal differentiation in the vertebrate lens.lengsin is an eye lens-specific member of the glutamine synthetase (gs) superfamily. lengsin has no gs activity, suggesting that it has a structural rather than catalytic role in lens. in situ hybridization and immunofluorescence showed that lengsin is expressed in terminally differentiating secondary lens fiber cells. yeast two-hybrid (y2h) and recombinant protein experiments showed that full-length lengsin can bind the 2b filament region of vimentin. in affinity chromatography, lengsin also bo ...200818178558
sirt1 acts as a nutrient-sensitive growth suppressor and its loss is associated with increased ampk and telomerase activity.sirt1, the mammalian homolog of sir2 in saccharomyces cerevisiae, is an nad-dependent deacetylase implicated in regulation of lifespan. by designing effective short hairpin rnas and a silent shrna-resistant mutant sirt1 in a genetically defined system, we show that efficient inhibition of sirt1 in telomerase-immortalized human cells enhanced cell growth under normal and nutrient limiting conditions. hematopoietic stem cells obtained from sirt1-deficient mice also showed increased growth capacity ...200818184747
prohibitin-1 maintains the angiogenic capacity of endothelial cells by regulating mitochondrial function and senescence.prohibitin 1 (phb1) is a highly conserved protein that is mainly localized to the inner mitochondrial membrane and has been implicated in regulating mitochondrial function in yeast. because mitochondria are emerging as an important regulator of vascular homeostasis, we examined phb1 function in endothelial cells. phb1 is highly expressed in the vascular system and knockdown of phb1 in endothelial cells increases mitochondrial production of reactive oxygen species via inhibition of complex i, whi ...200818195103
slic-1/sorting nexin 20: a novel sorting nexin that directs subcellular distribution of psgl-1.p-selectin glycoprotein ligand-1 (psgl-1) is a mucin-like glycoprotein expressed on the surface of leukocytes that serves as the major ligand for the selectin family of adhesion molecules and functions in leukocyte tethering and rolling on activated endothelium and platelets. previous studies have implicated the highly conserved cytoplasmic domain of psgl-1 in regulating outside-in signaling of integrin activation. however, molecules that physically and functionally interact with this domain are ...200818196517
comparative transcriptional profiling and evolutionary analysis of the gram domain family in eukaryotes.the gram domain was found in glucosyltransferases, myotubularins and other membrane-associated proteins. so far, functions for majority of these proteins are yet to be uncovered. in order to address the evolutionary and functional significance of this family members, we have performed a comprehensive investigation on their genome-wide identification, phylogenetic relationship and expression divergence in five different organisms representing monocot/dicot plants, vertebrate/invertebrate animals ...200818201690
detection and identification of plasma proteins that bind glialcam using proteinchip arrays, seldi-tof ms, and nano-lc ms/ms.in order to fully understand biological processes it is essential to identify interactions in protein complexes. there are several techniques available to study this type of interactions, such as yeast two-hybrid screens, affinity chromatography, and coimmunoprecipitation. we propose a novel strategy to identify protein-protein interactions, comprised of first detecting the interactions using proteinchips and seldi-tof ms, followed by the isolation of the interacting proteins through affinity be ...200818203261
functional trends in structural classes of the dna binding domains of regulatory transcription factors.the dna-binding domain (dbd) structure of a regulatory transcription factor (tf) is important in determining its dna sequence specificity, but it is unclear whether a relationship exists between dbd structure and general tf biological function or regulatory mechanism. we observed moderate enrichment of functional annotation terms among tfs of the same structural class in escherichia coli, saccharomyces cerevisiae, drosophila melanogaster, or mus musculus, suggesting some preference for tfs of si ...200818229706
high mobility group box (hmgb) proteins of plasmodium falciparum: dna binding proteins with pro-inflammatory activity.high mobility group box chromosomal protein 1 (hmgb1), known as an abundant, non-histone architectural chromosomal protein, is highly conserved across different species. homologues of hmgb1 were identified and cloned from malaria parasite, plasmodium falciparum. sequence analyses showed that the p. falciparum hmgb1 (pfhmgb1) exhibits 45, 23 and 18%, while pfhmgb2 shares 42, 21 and 17% homology with saccharomyces cerevisiae, human and mouse hmg box proteins respectively. parasite pfhmgb1and pfhmg ...200818234548
identification of a novel leucine-rich repeat protein and candidate pp1 regulatory subunit expressed in developing spermatids.spermatogenesis is comprised of a series of highly regulated developmental changes that transform the precursor germ cell into a highly specialized spermatozoon. the last phase of spermatogenesis, termed spermiogenesis, involves dramatic morphological change including formation of the acrosome, elongation and condensation of the nucleus, formation of the flagella, and disposal of unnecessary cytoplasm. a prominent cytoskeletal component of the developing spermatid is the manchette, a unique micr ...200818237440
a novel mammalian trans-membrane protein reveals an alternative initiation pathway for autophagy.autophagy is an early cellular event during acute pancreatitis, a disease defined as pancreas self-digestion. the vacuole membrane protein 1 (vmp1) is a trans-membrane protein highly activated in acinar cells early during pancreatitis-induced autophagy and it remains in the autophagosomal membrane. we have shown that vmp1 expression is able to trigger autophagy in mammalian cells, even under nutrient-replete conditions. vmp1 is induced by autophagy stimuli and its expression is required for auto ...200818253086
reversal of aging by nfkappab blockade.genetic studies in model organisms such as yeast, worms, flies, and mice leading to lifespan extension suggest that longevity is subject to regulation. in addition, various system-wide interventions in old animals can reverse features of aging. to better understand these processes, much effort has been put into the study of aging on a molecular level. in particular, genome-wide microarray analysis of differently aged individual organisms or tissues has been used to track the global expression ch ...200818256548
pi3k signaling of autophagy is required for starvation tolerance and virulenceof cryptococcus neoformans.autophagy is a process by which cells recycle cytoplasm and defective organelles during stress situations such as nutrient starvation. it can also be used by host cells as an immune defense mechanism to eliminate infectious pathogens. here we describe the use of autophagy as a survival mechanism and virulence-associated trait by the human fungal pathogen cryptococcus neoformans. we report that a mutant form of c. neoformans lacking the vps34 pi3k (vps34delta), which is known to be involved in au ...200818259613
a genetic screen for saccharomyces cerevisiae mutants affecting proteasome function, using a ubiquitin-independent substrate.the great majority of proteasome substrates are marked for degradation by the attachment of polyubiquitin chains. ornithine decarboxylase is degraded by the proteasome in the absence of this modification. we previously showed that this mechanism of degradation was conserved in eukaryotic cells. here we use a reporter destabilized by mouse ornithine decarboxylase to screen non-essential saccharomyces cerevisiae deletion mutants. we identified novel mutants that affect both ubiquitin-dependent and ...200818260085
dual compartmental localization and function of mammalian nadp+-specific isocitrate dehydrogenase in yeast.isozymes of nadp+-specific isocitrate dehydrogenase (idp) provide nadph in cytosolic, mitochondrial, and peroxisomal compartments of eukaryotic cells. analyses of purified idp isozymes from yeast and from mouse suggest a general correspondence of ph optima for catalysis and pi values with ph values reported for resident cellular compartments. however, mouse idp2, which partitions between cytosolic and peroxisomal compartments in mammalian cells, exhibits a broad ph optimum and an intermediate pi ...200818275837
trim11 increases expression of dopamine beta-hydroxylase gene by interacting with phox2b.the homeodomain transcription factor phox2b is one of the key determinants involved in the development of noradrenergic (na) neurons in both the central nervous system (cns) and the peripheral nervous system (pns). using yeast two-hybrid screening, we isolated a phox2b interacting protein, trim11, which belongs to trim (tripartite motif) or rbcc proteins family, and contains a ring domain, b-boxes, a coiled-coil domain, and the b30.2/spry domain. protein-protein interaction assays showed that ph ...200818275850
structural and biochemical characterization of the krlb region in insulin receptor substrate-2.insulin receptor substrates 1 and 2 (irs1 and -2) are crucial adaptor proteins in mediating the metabolic and mitogenic effects of insulin and insulin-like growth factor 1. these proteins consist of a pleckstrin homology domain, a phosphotyrosine binding domain and a c-terminal region containing numerous sites of tyrosine, serine and threonine phosphorylation. previous yeast two-hybrid studies identified a region unique to irs2, termed the kinase regulatory-loop binding (krlb) region, which inte ...200818278056
baf60a interacts with p53 to recruit the swi/snf complex.to understand the tumor-suppressing mechanism of the swi/snf chromatin remodeling complex, we investigated its molecular relationship with p53. using the prep4-luc episomal reporter, we first demonstrated that p53 utilizes the chromatin remodeling activity of the swi/snf complex to initiate transcription from the chromatin-structured promoter. among the components of the swi/snf complex, we identified baf60a as a mediator of the interaction with p53 by the yeast two-hybrid assay. p53 directly in ...200818303029
deoxynucleic acids from cryptococcus neoformans activate myeloid dendritic cells via a tlr9-dependent pathway.the mechanism of host cell recognition of cryptococcus neoformans, an opportunistic fungal pathogen in immunocompromised patients, remains poorly understood. in the present study, we asked whether the dna of this yeast activates mouse bone marrow-derived myeloid dendritic cells (bm-dcs). bm-dcs released il-12p40 and expressed cd40 upon stimulation with cryptococcal dna, and the response was abolished by treatment with dnase, but not with rnase. il-12p40 production and cd40 expression were attenu ...200818322216
novel oxidative stress-responsive gene ers25 functions as a regulator of the heat-shock and cell death response.members of the yeast p24 family, including emp24p and erv25p, exist as heteromeric complexes that have been proposed to cycle between the endoplasmic reticulum (er) and golgi compartments. the specific functions and sites of action of p24 proteins are still unknown. here we identified a human homolog of the yeast p24 family of proteins, named ers25 (endoplasmic reticulum stress-response protein 25), and investigated its role in stress response. ers25 is predicted to have an er localization signa ...200818326488
distinct roles of xrcc4 and ku80 in non-homologous end-joining of endonuclease- and ionizing radiation-induced dna double-strand breaks.non-homologous end-joining (nhej) of dna double-strand breaks (dsbs) is mediated by two protein complexes comprising ku80/ku70/dna-pkcs/artemis and xrcc4/ligaseiv/xlf. loss of ku or xrcc4/ligaseiv function compromises the rejoining of radiation-induced dsbs and leads to defective v(d)j recombination. in this study, we sought to define how xrcc4 and ku80 affect nhej of site-directed chromosomal dsbs in murine fibroblasts. we employed a recently developed reporter system based on the rejoining of ...200818332040
bin3 deletion causes cataracts and increased susceptibility to lymphoma during aging.bin3 encodes an evolutionarily conserved and ubiquitously expressed member of the bar superfamily of curved membrane and gtpase-binding proteins, which includes the bar, pch/f-bar, and i-bar adapter proteins implicated in signal transduction and vesicular trafficking. in humans, bin3 maps to chromosome 8p21.3, a region widely implicated in cancer suppression that is often deleted in non-hodgkin's lymphomas and various epithelial tumors. yeast studies have suggested roles for this gene in filamen ...200818339847
cryptococcal xylosyltransferase 1 (cxt1p) from cryptococcus neoformans plays a direct role in the synthesis of capsule polysaccharides.the opportunistic yeast cryptococcus neoformans causes serious disease in humans and expresses a prominent polysaccharide capsule that is required for its virulence. little is known about how this capsule is synthesized. we previously identified a beta1,2-xylosyltransferase (cxt1p) with in vitro enzymatic activity appropriate for involvement in capsule synthesis. here, we investigate c. neoformans strains in which the corresponding gene has been deleted (cxt1delta). loss of cxt1 does not affect ...200818347023
evolutionary and transcriptional analysis of karyopherin beta superfamily proteins.in eukaryotes, karyopherin beta superfamily proteins mediate nucleocytoplasmic transport of macromolecules. we investigated the evolutionary and transcriptional patterns of these proteins using bioinformatics approaches. no obvious homologs were found in prokaryotes, but an extensive set of beta-karyopherin proteins was found in yeast. among 14 beta-karyopherins of saccharomyces cerevisiae, eight corresponded to their human orthologs directly without diversification, two were lost, and the remai ...200818353765
identification of zag1, a novel protein expressed in mouse preimplantation, and its putative roles in zygotic genome activation.we isolated a mouse cdna, zag1 (zygotic gene activation-associated gene 1), that has an open reading frame of 1,728-bp encoding a protein of 66.2 kda including both a bipartite nuclear targeting sequence and a p-loop motif containing nucleoside triphosphate hydrolase motifs. northern blot analysis of mouse tissues showed that zag1 was widely expressed but was especially prominent in the ovary and testis. rt-pcr analysis of in vitro fertilized embryos showed that the abundance of zag1 transcripts ...200818360097
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