| a lectin hplc method to enrich selectively-glycosylated peptides from complex biological samples. | glycans are an important class of post-translational modifications. typically found on secreted and extracellular molecules, glycan structures signal the internal status of the cell. glycans on tumor cells tend to have abundant sialic acid and fucose moieties. we propose that these cancer-associated glycan variants be exploited for biomarker development aimed at diagnosing early-stage disease. accordingly, we developed a mass spectrometry-based workflow that incorporates chromatography on affini ... | 2009 | 19798022 |
| identification of salivary components that induce transition of hyphae to yeast in candida albicans. | candida albicans, the major human fungal pathogen, undergoes a reversible morphological transition from single yeast cells to pseudohyphae and hyphae filaments. the hyphae form is considered the most invasive form of the fungus. the purpose of this study is to investigate the effect of saliva on hyphae growth of c. albicans. candida albicans hyphae were inoculated in roswell park memorial institute medium with whole saliva, parotid saliva or buffer mimicking the saliva ion composition, and cultu ... | 2009 | 19799638 |
| the natural product aristolactam aiiia as a new ligand targeting the polo-box domain of polo-like kinase 1 potently inhibits cancer cell proliferation. | to search for novel inhibitors of human polo-like kinase 1 (plk1), which plays important roles in various aspects of mitotic progression and is believed as a promising anti-cancer drug target, and further investigate the potential inhibition mechanism of active compounds against plk1, thus developing potent anti-tumor lead compounds. | 2009 | 19801998 |
| mammalian glutaminyl cyclases and their isoenzymes have identical enzymatic characteristics. | glutaminyl cyclases (qcs) catalyze the formation of pyroglutamate residues at the n-terminus of several peptides and proteins from plants and animals. recently, isoenzymes of mammalian qcs have been identified. in order to gain further insight into the biochemical characteristics of isoqcs, the human and murine enzymes were expressed in the secretory pathway of pichia pastoris. replacement of the n-terminal signal anchor by an alpha-factor prepropeptide from saccharomyces cerevisiae resulted in ... | 2009 | 19804409 |
| nbs1 flexibly tethers ctp1 and mre11-rad50 to coordinate dna double-strand break processing and repair. | the nijmegen breakage syndrome 1 (nbs1) subunit of the mre11-rad50-nbs1 (mrn) complex protects genome integrity by coordinating double-strand break (dsb) repair and checkpoint signaling through undefined interactions with atm, mdc1, and sae2/ctp1/ctip. here, fission yeast and human nbs1 structures defined by x-ray crystallography and small angle x-ray scattering (saxs) reveal nbs1 cardinal features: fused, extended, fha-brct(1)-brct(2) domains flexibly linked to c-terminal mre11- and atm-binding ... | 2009 | 19804755 |
| rational extension of the ribosome biogenesis pathway using network-guided genetics. | biogenesis of ribosomes is an essential cellular process conserved across all eukaryotes and is known to require >170 genes for the assembly, modification, and trafficking of ribosome components through multiple cellular compartments. despite intensive study, this pathway likely involves many additional genes. here, we employ network-guided genetics-an approach for associating candidate genes with biological processes that capitalizes on recent advances in functional genomic and proteomic studie ... | 2009 | 19806183 |
| threshold selection in gene co-expression networks using spectral graph theory techniques. | gene co-expression networks are often constructed by computing some measure of similarity between expression levels of gene transcripts and subsequently applying a high-pass filter to remove all but the most likely biologically-significant relationships. the selection of this expression threshold necessarily has a significant effect on any conclusions derived from the resulting network. many approaches have been taken to choose an appropriate threshold, among them computing levels of statistical ... | 2009 | 19811688 |
| complement receptor 3, not dectin-1, is the major receptor on human neutrophils for beta-glucan-bearing particles. | we investigated the role of the beta-glucan receptor, dectin-1, in the response of human neutrophils to unopsonized saccharomyces cerevisiae and its major beta-glucan-containing capsular constituent, zymosan. although reported to be indispensable for yeast phagocytosis in murine phagocytes, human dectin-1 was not involved in the phagocytosis of s. cerevisiae or zymosan by human neutrophils. phagocytosis of yeast particles proved to be completely dependent on cd11b/cd18, also known as complement ... | 2009 | 19811837 |
| replication protein a (rpa1a) is required for meiotic and somatic dna repair but is dispensable for dna replication and homologous recombination in rice. | replication protein a (rpa), a highly conserved single-stranded dna-binding protein in eukaryotes, is a stable complex comprising three subunits termed rpa1, rpa2, and rpa3. rpa is required for multiple processes in dna metabolism such as replication, repair, and homologous recombination in yeast (saccharomyces cerevisiae) and human. most eukaryotic organisms, including fungi, insects, and vertebrates, have only a single rpa gene that encodes each rpa subunit. arabidopsis (arabidopsis thaliana) ... | 2009 | 19812186 |
| cryptococcal disease in patients with or without human immunodeficiency virus: clinical presentation and monitoring of serum cryptococcal antigen titers. | cryptococcus neoformans is an encapsulated pathogenic yeast that causes a wide range of clinical manifestations. the serum cryptococcal latex agglutination test is a simple, rapid, and reliable diagnostic test for cryptococcosis. this study was performed to assess the clinical relevance of serum cryptococcal antigen (crag) titer in patients with cryptococcosis with or without human immunodeficiency virus (hiv). | 2009 | 19812855 |
| [progresses in recombinant human serum albumin]. | human serum albumin (hsa) is an important biological product in clinical pharmacy practice at present. its main clinical use is in maintaining colloid osmotic pressure and increasing circulating plasma volume so as to cure hemorrhagic shock, burn, cancer, hypercytosis, hypoalbuminosis, etc. hsa is isolated by fractionating human plasma, which entails possible contamination by viruses or prions. recombinant human serum albumin (rhsa) has been successfully produced by biological engineering. the s ... | 2009 | 19813636 |
| efficient expression and secretion of recombinant human growth hormone in the methylotrophic yeast pichia pastoris: potential applications for other proteins. | a simple high yielding process for the production of rhgh (recombinant human growth hormone) in the pichia pastoris system is described. the approach adopted the addition of surfactants during fermentation along with methanol induction. a pichia integrant harbouring multiple-copy, non-codon-optimized hgh showed poor expression in complex and defined media. inclusion of the surfactants tween 20 or tween 80 during induction enhanced the expression levels significantly in shake flask studies. the c ... | 2009 | 19814714 |
| miz1 is a signal- and pathway-specific modulator or regulator (smor) that suppresses tnf-alpha-induced jnk1 activation. | the proinflammatory cytokine tnf-alpha exerts its pleiotropic functions through activation of multiple downstream effectors, including jnk1. yet, the underlying regulatory mechanism is incompletely understood. here, we report that the transcription factor myc-interacting zinc-finger protein 1 (miz1) selectively suppresses tnf-alpha-induced jnk1 activation and cell death independently of its transcription activity. proteomics analysis and yeast two-hybrid screening reveal that miz1 is a jnk-assoc ... | 2009 | 19815509 |
| identification of the n-glycosylation sites on recombinant bovine cd38 expressed in pichia pastoris: their impact on enzyme stability and catalytic activity. | bovine cd38, a type ii glycoprotein, contains two potential n-glycosylation sites (asn-201 and asn-268) in its extracellular domain. this contrasts with the other mammalian members of the adp-ribosyl cyclase family, such as human cd38 and bst-1/cd157, in which four such sites are present. our study was designed to determine the occupancy of these sites in a recombinant form of this ecto-enzyme and to evaluate its impact on the protein stability and catalytic functions. to that end we have succes ... | 2010 | 19818407 |
| budding yeast dbf4 sequences required for cdc7 kinase activation and identification of a functional relationship between the dbf4 and rev1 brct domains. | cdc7-dbf4 is a two-subunit kinase required for initiating dna replication. the dbf4 regulatory subunit is required for cdc7 kinase activity. previous studies have shown that the c termini of dbf4 orthologs encode a single (putative) c(2)h(2) zinc (zn) finger, referred to as "motif c." by mutational analysis we show that the zn finger is not required for the essential function of dbf4. however, deletion and point mutants altering conserved zn-finger residues exhibit a substantially slowed s-phase ... | 2009 | 19822727 |
| cdh1 controls the stability of tacc3. | transforming acidic coiled-coil protein 3 (tacc3) was reported to be important for regulating mitotic spindle assembly and chromosome segregation. while the protein level of tacc3 was shown to be altered during cell cycle progression, the molecular mechanism in controlling tacc3 level is unclear. here, we show that tacc3 protein level can be regulated by cdh1, a well known activator of anaphase-promoting complex/cyclosome. we identified cdh1 as an interacting partner of tacc3 by a yeast array sc ... | 2009 | 19823035 |
| analysis of candida albicans plasma membrane proteome. | the opportunistic human fungal pathogen candida albicans causes a wide variety of infections including deep systemic syndromes. the c. albicans plasma membrane is an important interface in the host-pathogen relationship. the plasma membrane proteins mediate a variety of functions, including sensing and signalling to the external environment, in which the glycosylphosphatidylinositol (gpi)-anchored membrane proteins play a crucial role. a subproteomic approach to obtain a global picture of the pr ... | 2009 | 19824013 |
| high-throughput cloning of human liver complete open reading frames using homologous recombination in escherichia coli. | in this article, we describe a high-throughput cloning method, seamless enzyme-free cloning (sefc), which allows one-step assembly of dna fragments in vivo via homologous recombination in escherichia coli. in the method, the desired open reading frame (orf) is amplified by use of orf-specific primers with flanking sequences identical to the two ends of a linearized vector. the polymerase chain reaction (pcr) product and the linearized vector are then cotransformed into e. coli cells, where the o ... | 2010 | 19835833 |
| characterization of papillomavirus e1 helicase mutants defective for interaction with the sumo-conjugating enzyme ubc9. | the e1 helicase from bpv and hpv16 interacts with ubc9 to facilitate viral genome replication. we report that hpv11 e1 also interacts with ubc9 in vitro and in the yeast two-hybrid system. residues in e1 involved in oligomerization (353-435) were sufficient for binding to ubc9 in vitro, but the origin-binding and atpase domains were additionally required in yeast. nuclear accumulation of bpv e1 was shown previously to depend on its interaction with ubc9 and sumoylation on lysine 514. in contrast ... | 2009 | 19836047 |
| n- and o-linked oligosaccharides completely lack galactose residues in the gms1och1 mutant of schizosaccharomyces pombe. | unlike their counterparts in budding yeast saccharomyces cerevisiae, the glycoproteins of schizosaccharomyces pombe contain, in addition to alpha-d-mannose (man), a large number of alpha-d-galactose (gal) residues. in both yeasts, large outer chains are attached to the oligosaccharide cores of glycoproteins during export via golgi. formation of the yeast-specific large outer chain is initiated by alpha-1,6-mannosylatransferase encoded by the och1+ gene, the disruption of which blocked outer chai ... | 2010 | 19844703 |
| determination of echinocandin mics for candida species in less than 8 hours: comparison of the rapid susceptibility assay with the clinical and laboratory standards institute's broth microdilution assay. | the echinocandins prevent fungal cell wall synthesis by inhibiting beta-1,3-glucan synthesis, a significant glucose-consuming process. previous studies suggested that echinocandin inhibitory activity is evident within 1 h of exposure. we hypothesized that a susceptibility assay based on glucose consumption may provide clinically useful mics rapidly. the rapid susceptibility assay (rsa), which provides mics in less than 8 h, was compared with the standard broth microdilution susceptibility assay ... | 2009 | 19846655 |
| the role of sex in fungal evolution. | sex in fungi is often associated with dispersal and dormancy, but in many species is not required for reproduction, and its evolutionary genetic role is unclear. sex can accelerate adaptation to a new environment, and recombination in saccharomyces cerevisiae, though historically rare, has had a prominent role in the origins of many sequenced strains, in particular the origin of clinical strains from domesticated ancestors. sex and recombination have recently been discovered in several human pat ... | 2009 | 19853499 |
| conserved telomere maintenance component 1 interacts with stn1 and maintains chromosome ends in higher eukaryotes. | orthologs of the yeast telomere protein stn1 are present in plants, but other components of the cdc13/stn1/ten1 (cst) complex have only been found in fungi. here we report the identification of conserved telomere maintenance component 1 (ctc1) in plants and vertebrates. ctc1 encodes an approximately 140 kda telomere-associated protein predicted to contain multiple ob-fold domains. arabidopsis mutants null for ctc1 display a severe telomere deprotection phenotype accompanied by a rapid onset of d ... | 2009 | 19854131 |
| inhibition of hepatic carnitine palmitoyl-transferase i (cpt ia) by valproyl-coa as a possible mechanism of valproate-induced steatosis. | carnitine palmitoyl-transferase i (cpt i) catalyses the synthesis of long-chain (lc)-acylcarnitines from lc-acyl-coa esters. it is the rate-limiting enzyme of mitochondrial fatty acid beta-oxidation (fao) pathway and its activity is regulated by malonyl-coa. the antiepileptic drug valproic acid (vpa) is a branched chain fatty acid that is activated to the respective coa ester in the intra- and extra-mitochondrial compartments. this drug has been associated with a clear inhibition of mitochondria ... | 2010 | 19854160 |
| physical and genetic interactions of yeast cwc21p, an ortholog of human srm300/srrm2, suggest a role at the catalytic center of the spliceosome. | in saccharomyces cerevisiae, cwc21p is a protein of unknown function that is associated with the nineteen complex (ntc), a group of proteins involved in activating the spliceosome to promote the pre-mrna splicing reaction. here, we show that cwc21p binds directly to two key splicing factors-namely, prp8p and snu114p-and becomes the first ntc-related protein known to dock directly to u5 snrnp proteins. using a combination of proteomic techniques we show that the n-terminus of prp8p contains an in ... | 2009 | 19854871 |
| dh166, a beta-carboline derivative, inhibits the kinase activity of plk1. | a better way to treat complex diseases such as cancer is to aim for several targets at once. beta-carboline derivatives have been shown to have anticancer activity, but these compounds may target several enzymes required for cell division. polo-like kinases (plks) are well conserved serine/threonine kinases and plk1 plays multiple roles in cell proliferation. thus, plk1 is one of the attractive mitotic targets for anticancer drugs. we found that dh166, a beta-carboline derivative, inhibits the g ... | 2009 | 19855194 |
| the c3-activator system: an alternate pathway of complement activation. | evidence has accumulated indicating the existence of a second complement activation mechanism which is functionally analogous to c1, c2, and c4. the noncomplement protein c3pa, previously recognized through its ability to form a complex enzyme with a protein from cobra venom, appears to be the precursor of the c4,2 analogue. it is a thermolabile beta-globulin with a molecular weight of 80,000. when serum is treated with naturally occurring plant or bacterial polysaccharides, the c3pa is cleaved ... | 1971 | 19867385 |
| terbinafine: a pharmacological and clinical review. | terbinafine (trb) is an allylamine antifungal agent that has been available for more than a decade. it is now used for the treatment of dermatophytic infections and onychomycosis. despite several studies having demonstrated the efficacy of terbinafine against nondermatophytic infections including azole-resistant candidiasis, invasive aspergillosis, disseminated fusariosis and scedosporiosis, the role of trb in the management of these infections remains greatly underappreciated. a brief review of ... | 2009 | 19874252 |
| versatile assays for high throughput screening for activators or inhibitors of intracellular proteases and their cellular regulators. | intracellular proteases constitute a class of promising drug discovery targets. methods for high throughput screening against these targets are generally limited to in vitro biochemical assays that can suffer many technical limitations, as well as failing to capture the biological context of proteases within the cellular pathways that lead to their activation. methods #entitystartx00026; | 2009 | 19876397 |
| the yeast candida albicans evades human complement attack by secretion of aspartic proteases. | candida albicans, which represents one of the most important human pathogenic yeasts, is directly attacked by the host innate immune system upon infection. however this pathogen has developed multiple strategies to escape host immune defense. here, we show that c. albicans secreted proteases interfere and inactivate host innate immune effector components, such as complement proteins. secreted aspartic proteases (saps) in the culture supernatant of c. albicans cells and also recombinant sap1, sap ... | 2009 | 19880183 |
| serms and sarms: detection of their activities with yeast based bioassays. | selective estrogen receptor modulators (serms) and selective androgen receptor modulators (sarms) are compounds that activate their cognate receptor in particular target tissues without affecting other organs. many of these compounds will find their use in therapeutic treatments. however, they also will have a high potential for misuse in veterinary practice and the sporting world. here we demonstrate that yeast estrogen and androgen bioassays can be used to detect serms and sarms, and are also ... | 2010 | 19883760 |
| diagnostic pcr tests for microsporum audouinii, m. canis and trichophyton infections. | since traditional diagnosis of dermatophyte infections is slow, we present a rapid new pcr test for detection of trichophyton spp., microsporum canis and m. audouinii infections. the performance of the test was evaluated with: 58 dermatophyte isolates; 10 yeast, mould and human dna control samples; 25 routine specimens from patients suspected of having dermatophytosis; 10 hair specimens from guinea pigs experimentally infected with m. canis; and two samples from un-infected control animals. dna ... | 2010 | 19886764 |
| molecular characterisation of mek1/2- and mkk3/6-like mitogen-activated protein kinase kinases (mapkk) from the fox tapeworm echinococcus multilocularis. | mitogen-activated protein kinase kinases (mapkks) are essential components of evolutionary conserved signalling modules that regulate a variety of fundamental cellular processes in response to environmental stimuli. to date, no mapkk ortholog has been characterised in free-living or parasitic flatworm species. here, we report the identification and molecular characterisation of two such molecules in the human parasitic cestode echinococcus multilocularis, the causative agent of alveolar echinoco ... | 2010 | 19887070 |
| structure and function of the plaa/ufd3-p97/cdc48 complex. | plaa (ortholog of yeast doa1/ufd3, also know as human plap or phospholipase a2-activating protein) has been implicated in a variety of disparate biological processes that involve the ubiquitin system. it is linked to the maintenance of ubiquitin levels, but the mechanism by which it accomplishes this is unclear. the c-terminal pul (plap, ufd3p, and lub1p) domain of plaa binds p97, an aaa atpase, which among other functions helps transfer ubiquitinated proteins to the proteasome for degradation. ... | 2010 | 19887378 |
| nutrient sensing g protein-coupled receptors: interesting targets for antifungals? | g protein-coupled receptors (gpcrs) are important targets for various drugs that are on the market. an important area in which we urgently need novel drug targets is the field of antifungals. recently a new class of nutrient sensing g protein-coupled receptors have been identified in fungi. the founding member of this novel class of gpcrs is gpr1, the sucrose/glucose sensing receptor of saccharomyces cerevisiae. this receptor activates the camp-pka pathway that is important for the yeast-to-pseu ... | 2009 | 19888799 |
| tlr-2, tlr-4 and dectin-1 expression in human monocytes and neutrophils stimulated by paracoccidioides brasiliensis. | paracoccidioidomycosis (pcm) is an endemic mycosis in latin america caused by the dimorphic fungus paracoccidioides brasiliensis. the pattern of the immune responses to p. brasiliensis determines the disease progression and clinical outcome. innate immune response is mediated by phagocytic cells, such as macrophage and neutrophils, which ingest and kill invading pathogens and then trigger the adaptive immune system through the secretion of cytokines and chemokines. the c-type like lectin recepto ... | 2009 | 19888805 |
| human sec3 protein is a novel transcriptional and translational repressor of flavivirus. | the flaviviridae family consists of several medically important pathogens such as west nile virus (wnv) and dengue virus (denv). flavivirus capsid (c) protein is a key structural component of virus particles. however, the role of c protein in the pathogenesis of arthropod-borne flaviviruses is poorly understood. to examine whether flavivirus c protein can associate with cellular proteins, and contribute to viral pathogenesis, wnv/denv c protein was screened against a human brain/liver cdna yeast ... | 2010 | 19889084 |
| creating protein affinity reagents by combining peptide ligands on synthetic dna scaffolds. | a full understanding of the proteome will require ligands to all of the proteins encoded by genomes. while antibodies represent the principle affinity reagents used to bind proteins, their limitations have created a need for new ligands to large numbers of proteins. here we propose a general concept to obtain protein affinity reagents that avoids animal immunization and iterative selection steps. central to this process is the idea that small peptide libraries contain sequences that will bind to ... | 2009 | 19894711 |
| production of immunogenic human papillomavirus-16 major capsid protein derived virus like particles. | recombinant dna technology allows expression of the human papillomavirus (hpv) major capsid protein (l1) in heterologous expression systems and the recombinant protein self assembles to virus-like particles (vlp). we took up this study to produce recombinant hpv-16 l1 in yeast, establish the process of recombinant l1 derived vlp preparation and develop an elisa using vlp as the antigen for serological evaluation of anti hpv-16 l1 antibody status. | 2009 | 19901429 |
| arf antagonizes the ability of miz-1 to inhibit p53-mediated transactivation. | although myc-interacting zinc-finger protein-1 (miz-1) is known to be a poxvirus and zinc-finger (poz) transcription factor required for myc transcriptional repression, additional regulatory function of miz-1 is less well understood. using a yeast two-hybrid screen, we identified human alternate reading frame (arf) protein as a novel interaction partner of miz-1. the zinc-finger domain of miz-1 is involved in its binding to arf. in addition, we found that miz-1 was able to interact with p53 thro ... | 2010 | 19901969 |
| concordant regulation of translation and mrna abundance for hundreds of targets of a human microrna. | micrornas (mirnas) regulate gene expression posttranscriptionally by interfering with a target mrna's translation, stability, or both. we sought to dissect the respective contributions of translational inhibition and mrna decay to microrna regulation. we identified direct targets of a specific mirna, mir-124, by virtue of their association with argonaute proteins, core components of mirna effector complexes, in response to mir-124 transfection in human tissue culture cells. in parallel, we asses ... | 2009 | 19901979 |
| cohesin promotes the repair of ionizing radiation-induced dna double-strand breaks in replicated chromatin. | the cohesin protein complex holds sister chromatids together after synthesis until mitosis. it also contributes to post-replicative dna repair in yeast and higher eukaryotes and accumulates at sites of laser-induced damage in human cells. our goal was to determine whether the cohesin subunits smc1 and rad21 contribute to dna double-strand break repair in x-irradiated human cells in the g2 phase of the cell cycle. rna interference-mediated depletion of smc1 sensitized hela cells to x-rays. repair ... | 2010 | 19906707 |
| the human c-fos and tnfalpha au-rich elements show different effects on mrna abundance and protein expression depending on the reporter in the yeast pichia pastoris. | au-rich elements (ares) are located in the 3' untranslated region (3' utr) of their host genes and tightly regulate mrna degradation and expression. examples for this kind of regulation are the human proto-oncogene c-fos and the cytokine tnfalpha. despite large effort in this field, the exact mechanism of are-mediated mrna turnover remains unclear. in this work we analysed the effects of c-fos- and tnfalpha ares on mrna abundance and protein expression of selected human cdnas in the yeast pichia ... | 2010 | 19908204 |
| probing the metabolic aberrations underlying mutant huntingtin toxicity in yeast and assessing their degree of preservation in humans and mice. | metabolomics is a powerful multiparameter tool for evaluating phenotypic traits associated with disease processes. we have used (1)h nmr metabolome profiling to characterize metabolic aberrations in a yeast model of huntington's disease that are attributable to the mutant huntingtin protein's gain-of-toxic-function effects. a group of 11 metabolites (alanine, acetate, galactose, glutamine, glycerol, histidine, proline, succinate, threonine, trehalose, and valine) exhibited significant concentrat ... | 2010 | 19908918 |
| identification of novel pten-binding partners: pten interaction with fatty acid binding protein fabp4. | pten is a tumor suppressor with dual protein and lipid-phosphatase activity, which is frequently deleted or mutated in many human advanced cancers. recent studies have also demonstrated that pten is a promising target in type ii diabetes and obesity treatment. using c-terminal pten sequence in peg202-nls as bait, yeast two-hybrid screening on mouse embryo, colon cancer, and hela cdna libraries was carried out. isolated positive clones were validated by mating assay and identified through automat ... | 2010 | 19911253 |
| quantitative in vitro and in vivo characterization of the human p32t mutant itpase. | human itpase, encoded by the itpa gene, and its orthologs (rdgb in escherichia coli and ham1 in saccharomyces cerevisiae) exclude noncanonical nucleoside triphosphates (ntps) from ntp pools. deoxyinosine triphosphate (ditp) and 2'-deoxy-n-6-hydroxylaminopurine triphosphate are both hydrolyzed by itpase to yield the corresponding deoxynucleoside monophosphate and pyrophosphate. in addition, metabolites of thiopurine drugs such as azathioprine have been shown to be substrates for itpase. the itpa ... | 2010 | 19914375 |
| an auxin-based degron system for the rapid depletion of proteins in nonplant cells. | plants have evolved a unique system in which the plant hormone auxin directly induces rapid degradation of the aux/iaa family of transcription repressors by a specific form of the scf e3 ubiquitin ligase. other eukaryotes lack the auxin response but share the scf degradation pathway, allowing us to transplant the auxin-inducible degron (aid) system into nonplant cells and use a small molecule to conditionally control protein stability. the aid system allowed rapid and reversible degradation of t ... | 2009 | 19915560 |
| a genome-wide deletion mutant screen identifies pathways affected by nickel sulfate in saccharomyces cerevisiae. | the understanding of the biological function, regulation, and cellular interactions of the yeast genome and proteome, along with the high conservation in gene function found between yeast genes and their human homologues, has allowed for saccharomyces cerevisiae to be used as a model organism to deduce biological processes in human cells. here, we have completed a systematic screen of the entire set of 4,733 haploid s. cerevisiae gene deletion strains (the entire set of nonessential genes for th ... | 2009 | 19917080 |
| biochemical characterization of human ecdysoneless reveals a role in transcriptional regulation. | ecdysoneless (ecd) is an evolutionarily conserved protein and its function is essential for embryonic development in drosophila and cell growth in yeast. however, its function has remained unknown until recently. studies in yeast suggested a potential role of ecd in transcription; however, ecd lacks a dna-binding domain. using a gal4-luciferase reporter assay and a gal4 dna-binding domain fusion with ecd or its mutants, we present evidence that human ecd has a transactivation activity in its c-t ... | 2010 | 19919181 |
| cyp21-catalyzed production of the long-term urinary metandienone metabolite 17beta-hydroxymethyl-17 alpha-methyl-18-norandrosta-1,4,13-trien-3-one: a contribution to the fight against doping. | anabolic-androgenic steroids are some of the most frequently misused drugs in human sports. recently, a previously unknown urinary metabolite of metandienone, 17beta-hydroxymethyl-17 alpha-methyl-18-norandrosta-1,4,13-trien-3-one (20oh-normd), was discovered via lc-ms/ms and gc-ms. this metabolite was reported to be detected in urine samples up to 19 days after administration of metandienone. however, so far it was not possible to obtain purified reference material of this metabolite and to conf ... | 2010 | 19919183 |
| lc-ms/ms analysis of epoxyalcohols and epoxides of arachidonic acid and their oxygenation by recombinant cyp4f8 and cyp4f22. | cyp4f22 and cyp4f8 are expressed in epidermis, and mutations of cyp4f22 are associated with lamellar ichthyosis. epoxyalcohols (heets) and epoxides (eets) of 20:4n-6 appear to be important for the water permeability barrier of skin. our aim was to study the ms/ms spectra and fragmentation of these compounds and to determine whether they were oxidized by cyp4f22 or cyp4f8 expressed in yeast. heets were prepared from 15-hydroperoxyeicosatetraenoic acid (15-hpete), 12-hpete, and their [(2)h(8)]labe ... | 2010 | 19919823 |
| identification of cenp-v as a novel microtubule-associating molecule that activates src family kinases through sh3 domain interaction. | the activation mechanisms of src family kinases (sfks) involve the dissociation of the intramolecular interaction between the src homology (sh) 3 and kinase domain. this process is mediated by the intermolecular attack of outer ligands to the sh3 domain. by using a yeast two-hybrid screen, we isolated a relevant ligand involved in the activation mechanisms of sfks. this molecule was found to be identical to a recently recognized kinetochore protein--designated as centromere protein (cenp)-v--whi ... | 2009 | 19930468 |
| defining the pathogenesis of the human atp12p w94r mutation using a saccharomyces cerevisiae yeast model. | studies in yeast have shown that a deficiency in atp12p prevents assembly of the extrinsic domain (f(1)) of complex v and renders cells unable to make atp through oxidative phosphorylation. de meirleir et al. (de meirleir, l., seneca, s., lissens, w., de clercq, i., eyskens, f., gerlo, e., smet, j., and van coster, r. (2004) j. med. genet. 41, 120-124) have reported that a homozygous missense mutation in the gene for human atp12p (huatp12p), which replaces trp-94 with arg, was linked to the deat ... | 2010 | 19933271 |
| multiple determinants within iron-responsive elements dictate iron regulatory protein binding and regulatory hierarchy. | iron regulatory proteins (irps) are iron-regulated rna binding proteins that, along with iron-responsive elements (ires), control the translation of a diverse set of mrna with 5' ire. dysregulation of irp action causes disease with etiology that may reflect differential control of ire-containing mrna. ires are defined by a conserved stem-loop structure including a midstem bulge at c8 and a terminal cagugh sequence that forms an agu pseudo-triloop and n19 bulge. c8 and the pseudo-triloop nucleoti ... | 2010 | 19939970 |
| production of bioactive human alpha-defensin 5 in pichia pastoris. | human alpha-defensin 5 (hd5), a small cationic peptide, is expressed in paneth cell granules of small intestinal crypts. hd5 exhibits high antimicrobial activity against a broad spectrum of pathogenic agents, including bacteria, fungi, and viruses. in this study, the constitutive expression of hd5 antimicrobial peptide was achieved using the methylotrophic yeast, pichia pastoris (p. pastoris). hd5 cdna was amplified by polymerase chain reaction (pcr) using human lung cell cdna as template. the 9 ... | 2009 | 19940385 |
| a genome-wide rna interference screen identifies two novel components of the metazoan secretory pathway. | genetic screens in the yeast saccharomyces cerevisiae have identified many proteins involved in the secretory pathway, most of which have orthologues in higher eukaryotes. to investigate whether there are additional proteins that are required for secretion in metazoans but are absent from yeast, we used genome-wide rna interference (rnai) to look for genes required for secretion of recombinant luciferase from drosophila s2 cells. this identified two novel components of the secretory pathway that ... | 2010 | 19942856 |
| the novel peptide f29 facilitates the dna-binding ability of hypoxia-inducible factor-1alpha. | hypoxia-inducible factor-1alpha/beta (hif-1alpha/beta) is a heterodimeric transcriptional activator that mediates gene expression in response to hypoxia. hif-1alpha has been noted as an effective therapeutic target for ischemic diseases such as myocardiac infarction, stroke and cancer. by using a yeast two-hybrid system and a random peptide library, we found a 16-mer peptide named f29 that directly interacts with the bhlh-pas domain of hif-1alpha. we found that f29 facilitates the interaction of ... | 2009 | 19944015 |
| comparative deacetylase activity of wild type and mutants of sirt1. | sirt1, human ortholog of yeast sir2 protein, deacetylates histones and several other transcription factors. recently, sirt1 has emerged as a drug target for treating age related diseases, type ii diabetes, neurodegeneration, inflammation and cancer. here, we have optimized production of functionally active wild type full-length sirt1 protein and its n-terminal deleted mutants. in a comparative study, we found that the region containing 192-208 amino acids towards the n-terminus is critical for r ... | 2010 | 19944070 |
| abc transporters in saccharomyces cerevisiae and their interactors: new technology advances the biology of the abcc (mrp) subfamily. | members of the atp-binding cassette (abc) transporter superfamily exist in bacteria, fungi, plants, and animals and play key roles in the efflux of xenobiotic compounds, physiological substrates, and toxic intracellular metabolites. based on sequence relatedness, mammalian abc proteins have been divided into seven subfamilies, abc subfamily a (abca) to abcg. this review focuses on recent advances in our understanding of abc transporters in the model organism saccharomyces cerevisiae. we propose ... | 2009 | 19946134 |
| rack1 promotes breast carcinoma proliferation and invasion/metastasis in vitro and in vivo. | a yeast two-hybrid system was utilized to identify novel pi3k p110alpha-interacting proteins, of which receptor of activated protein kinase c1 (rack1) was chosen for successive detailed analyses. our aim was to investigate the function(s) of rack1 and its involvement in mechanisms of breast carcinoma proliferation and invasion/metastasis. experiments in breast carcinoma cell lines stably transfected with rack1, as well as nude mouse models, showed that rack1 promotes breast carcinoma proliferati ... | 2010 | 19946739 |
| identifying differentially expressed pathways via a mixed integer linear programming model. | the identification of genes and pathways involved in biological processes is a central problem in systems biology. recent microarray technologies and other high-throughput experiments provide information which sheds light on this problem. in this article, the authors propose a new computational method to detect active pathways, or identify differentially expressed pathways via integration of gene expression and interactomic data in a sophisticated and efficient manner. specifically, by using sig ... | 2009 | 19947773 |
| comparative study of human mitochondrial proteome reveals extensive protein subcellular relocalization after gene duplications. | gene and genome duplication is the principle creative force in evolution. recently, protein subcellular relocalization, or neolocalization was proposed as one of the mechanisms responsible for the retention of duplicated genes. this hypothesis received support from the analysis of yeast genomes, but has not been tested thoroughly on animal genomes. in order to evaluate the importance of subcellular relocalizations for retention of duplicated genes in animal genomes, we systematically analyzed nu ... | 2009 | 19948060 |
| mammalian rif1 contributes to replication stress survival and homology-directed repair. | rif1, originally recognized for its role at telomeres in budding yeast, has been implicated in a wide variety of cellular processes in mammals, including pluripotency of stem cells, response to double-strand breaks, and breast cancer development. as the molecular function of rif1 is not known, we examined the consequences of rif1 deficiency in mouse cells. rif1 deficiency leads to failure in embryonic development, and conditional deletion of rif1 from mouse embryo fibroblasts affects s-phase pro ... | 2009 | 19948482 |
| role of double-stranded dna translocase activity of human hltf in replication of damaged dna. | unrepaired dna lesions can block the progression of the replication fork, leading to genomic instability and cancer in higher-order eukaryotes. in saccharomyces cerevisiae, replication through dna lesions can be mediated by translesion synthesis dna polymerases, leading to error-free or error-prone damage bypass, or by rad5-mediated template switching to the sister chromatid that is inherently error free. while translesion synthesis pathways are highly conserved from yeast to humans, very little ... | 2010 | 19948885 |
| revealing human disease genes through analysis of the yeast mitochondrial proteome. | | 2009 | 19949294 |
| cloning, yeast expression, purification and biological activity of a truncated form of the soybean 7s globulin alpha' subunit involved in hep g2 cell cholesterol homeostasis. | a truncated form of alpha' chain (talpha'), the soybean 7s globulin subunit previously demonstrated to be active in controlling the cholesterol and triglyceride homeostasis in in vitro and in vivo models, was cloned and expressed in the yeast pichia pastoris. the recombinant polypeptide spanned 216 amino acid residues from the n-terminal side and included the n-terminal extension region of the soybean subunit. the talpha' polypeptide was purified by conventional biochemical techniques, and its p ... | 2010 | 19954942 |
| association of whirlin with cav1.3 (alpha1d) channels in photoreceptors, defining a novel member of the usher protein network. | usher syndrome is the most common form of hereditary deaf-blindness. it is both clinically and genetically heterogeneous. the ush2d protein whirlin interacts via its pdz domains with other usher-associated proteins containing a c-terminal type i pdz-binding motif. these proteins co-localize with whirlin at the region of the connecting cilium and at the synapse of photoreceptor cells. this study was undertaken to identify novel, usher syndrome-associated, interacting partners of whirlin and there ... | 2010 | 19959638 |
| single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays. | single plasmids designed to express the six human type i steroid hormone receptors and detect signaling activity are described in this report. these stably replicating plasmids reported ligand-induced transcriptional activation via lacz assays in baker's yeast (saccharomyces cerevisiae). the ligand concentrations needed to activate signaling in yeast expressing these plasmids spanned five orders of magnitude as based on comparisons of ec(50) values. radicicol, a direct inhibitor of heat shock pr ... | 2010 | 19962400 |
| network topological reordering revealing systemic patterns in yeast protein interaction networks. | identifying candidate genes/proteins involved in human disease specific molecular pathways or networks has been a primary focus of biomedical research. although node ranking and graph clustering methods can help identify localized topological properties in a network, it remains unclear how the results should be interpreted in biological functional context in systems-level. in complex biomolecular interaction networks, biomolecular entities may not have absolute ranks or clear cluster boundary am ... | 2009 | 19964716 |
| haploid genetic screens in human cells identify host factors used by pathogens. | loss-of-function genetic screens in model organisms have elucidated numerous biological processes, but the diploid genome of mammalian cells has precluded large-scale gene disruption. we used insertional mutagenesis to develop a screening method to generate null alleles in a human cell line haploid for all chromosomes except chromosome 8. using this approach, we identified host factors essential for infection with influenza and genes encoding important elements of the biosynthetic pathway of dip ... | 2009 | 19965467 |
| altered gene expression in the werner and bloom syndromes is associated with sequences having g-quadruplex forming potential. | the human werner and bloom syndromes (ws and bs) are caused by deficiencies in the wrn and blm recq helicases, respectively. wrn, blm and their saccharomyces cerevisiae homologue sgs1, are particularly active in vitro in unwinding g-quadruplex dna (g4-dna), a family of non-canonical nucleic acid structures formed by certain g-rich sequences. recently, mrna levels from loci containing potential g-quadruplex-forming sequences (pqs) were found to be preferentially altered in sgs1delta mutants, sugg ... | 2010 | 19966276 |
| yeast strains with n-terminally truncated ribosomal protein s5: implications for the evolution, structure and function of the rps5/rps7 proteins. | ribosomal protein (rp)s5 belongs to the family of the highly conserved rp's that contains rps7 from prokaryotes and rps5 from eukaryotes. alignment of rps5/rps7 from metazoans (homo sapiens), fungi (saccharomyces cerevisiae) and bacteria (escherichia coli) shows that the proteins contain a conserved central/c-terminal core region and possess variable n-terminal regions. yeast rps5 is 69 amino acids (aa) longer than the e. coli rps7 protein; and human rps5 is 48 aa longer than the rps7, respectiv ... | 2010 | 19969550 |
| the ras/camp/pka signaling pathway and virulence in candida albicans. | candidiasis is the most common cause of fungal infections, and the majority of these are caused by candida albicans. the protean pathogenic potential of c. albicans includes the capacity to infect diverse mucosal and epidermal surfaces as well as to disseminate via the bloodstream to internal organs, potentially causing system failure in cases of severe immunosuppression. many environmental niches in the host may be invaded by c. albicans through modulation of gene expression patterns while chan ... | 2009 | 19995187 |
| 2'-hydroxylation of genistein enhanced antioxidant and antiproliferative activities in mcf-7 human breast cancer cells. | bioconversion of the isoflavonoid genistein to 2'- hydroxygenistein (2'-hg) was performed using isoflavone 2'-hydroxylase (cyp81e1) heterologously expressed in yeast. a monohydroxylated product was analyzed by liquid chromatography-electrospray ionization-mass spectrometry (lc-esi-ms) and nmr spectrometry and was identified as 2'-hg. an initial bioconversion rate of 6% was increased up to 14% under optimized conditions. after recovery, the biological activity of 2'-hg was evaluated. bioconverted ... | 2009 | 19996686 |
| haspin: a newly discovered regulator of mitotic chromosome behavior. | the haspins are divergent members of the eukaryotic protein kinase family that are conserved in many eukaryotic lineages including animals, fungi, and plants. recently-solved crystal structures confirm that the kinase domain of human haspin has unusual structural features that stabilize a catalytically active conformation and create a distinctive substrate binding site. haspin localizes predominantly to chromosomes and phosphorylates histone h3 at threonine-3 during mitosis, particularly at inne ... | 2010 | 19997740 |
| prevalence of candida dubliniensis among cancer patients in kuwait: a 5-year retrospective study. | summary despite close genetic and phenotypic relationship of candida dubliniensis with candida albicans, its role in human disease is mostly restricted to oral colonisation, particularly among hiv-infected patients. the prevalence of c. dubliniensis in association with other disease conditions has been infrequently reported. in this study, we present data on the prevalence of c. dubliniensis among yeast species isolated from cancer patients over a 5-year period. a total of 1445 yeast isolates re ... | 2009 | 20002881 |
| the unstructured c-terminal tail of the 9-1-1 clamp subunit ddc1 activates mec1/atr via two distinct mechanisms. | dna damage checkpoint pathways operate to prevent cell-cycle progression in response to dna damage and replication stress. in s. cerevisiae, mec1-ddc2 (human atr-atrip) is the principal checkpoint protein kinase. biochemical studies have identified two factors, the 9-1-1 checkpoint clamp and the dpb11/topbp1 replication protein, as potential activators of mec1/atr. here, we show that g1 phase checkpoint activation of mec1 is achieved by the ddc1 subunit of 9-1-1, while dpb11 is dispensable. howe ... | 2009 | 20005839 |
| glucuronide production by whole-cell biotransformation using genetically engineered fission yeast schizosaccharomyces pombe. | drug metabolites generated by udp glycosyltransferases (ugts) are needed for drug development and toxicity studies, especially in the context of safety testing of metabolites during drug development. because chemical metabolite synthesis can be arduous, various biological approaches have been developed; however, no whole-cell biotransformation with recombinant microbes that express human ugts was yet achieved. in this study we expressed human udp glucose-6-dehydrogenase together with several hum ... | 2010 | 20008039 |
| factor h and factor h-related protein 1 bind to human neutrophils via complement receptor 3, mediate attachment to candida albicans, and enhance neutrophil antimicrobial activity. | the host complement system plays an important role in protection against infections. several human-pathogenic microbes were shown to acquire host complement regulators, such as factor h (cfh), that downregulate complement activation at the microbial surface and protect the pathogens from the opsonic and lytic effects of complement. because cfh can also bind to host cells, we addressed the role of cfh and cfh-related proteins as adhesion ligands in host-pathogen interactions. we show that the cfh ... | 2010 | 20008295 |
| nix is a selective autophagy receptor for mitochondrial clearance. | autophagy is the cellular homeostatic pathway that delivers large cytosolic materials for degradation in the lysosome. recent evidence indicates that autophagy mediates selective removal of protein aggregates, organelles and microbes in cells. yet, the specificity in targeting a particular substrate to the autophagy pathway remains poorly understood. here, we show that the mitochondrial protein nix is a selective autophagy receptor by binding to lc3/gabarap proteins, ubiquitin-like modifiers tha ... | 2010 | 20010802 |
| alternative splicing and gene polymorphism of the human tap3/sec14l4 gene. | three closely related human sec14p-like proteins (htap1, htap2, htap3, or sec14l2, sec14l3, sec14l4, respectively) have been described that are related to the saccharomyces cerevisiae sec14 protein. these proteins may participate in intracellular lipid transport and influence regulatory lipid-dependent events. here we report the isolation of an alternatively spliced htap3 cdna and a polymorphism within the coding region of the htap3/sec14l4 gene. | 2010 | 20012369 |
| function of topbp1 in genome stability. | human dna topoisomerase iibeta-binding protein 1 (topbp1) and its orthologues in other organisms are proteins consisting of multiple brct modules that have acquired several functions during evolution. these proteins execute their tasks by interacting with a great variety of proteins involved in nuclear processes. topbp1 is an essential protein that has numerous roles in the maintenance of the genomic integrity. in particular, it is required for the activation of atm and rad3-related (atr), a vit ... | 2010 | 20012580 |
| ire1alpha controls cyclin a1 expression and promotes cell proliferation through xbp-1. | ire1 is a conserved dual endoribonuclease/protein kinase that is indispensable for directing the endoplasmic reticulum (er) stress response in yeast, flies, and worms. in mammalian systems, however, the precise biological activities carried out by ire1alpha are unclear. here, molecular and chemical genetic approaches were used to control ire1 activity in a number of prostate cancer cell lines and the resulting impact on gene transcription, cell survival, and proliferation was examined. modulatin ... | 2010 | 20013084 |
| identification and characterization of the type 2c protein phosphatase ptc4p in the human fungal pathogen candida albicans. | type 2c protein phosphatases (pp2c) are monomeric enzymes and their activities require the presence of magnesium or manganese ions. there are seven pp2c genes, named from ptc1 to ptc7, in saccharomyces cerevisiae. in the current study we identified the captc4 gene in candida albicans and demonstrated that the captc4p protein is a typical pp2c enzyme, which is highly conserved in fungal species. deletion of captc4 renders candida cells sensitive to sodium and potassium ions as well as to antifung ... | 2010 | 20014041 |
| apolipoprotein e interacts with hepatitis c virus nonstructural protein 5a and determines assembly of infectious particles. | chronic hepatitis c virus (hcv) infection is a major cause of liver disease worldwide. restriction of hcv infection to human hepatocytes suggests that liver-specific host factors play a role in the viral life cycle. using a yeast-two-hybrid system, we identified apolipoprotein e (apoe) as a liver-derived host factor specifically interacting with hcv nonstructural protein 5a (ns5a) but not with other viral proteins. the relevance of apoe-ns5a interaction for viral infection was confirmed by co-im ... | 2010 | 20014138 |
| from meiosis to postmeiotic events: uncovering the molecular roles of the meiosis-specific recombinase dmc1. | in meiosis, the accurate segregation of maternal and paternal chromosomes is accomplished by homologous recombination. a central player in meiotic recombination is the dmc1 recombinase, a member of the reca/rad51 recombinase superfamily, which is widely conserved from viruses to humans. dmc1 is a meiosis-specific protein that functions with the ubiquitously expressed homolog, the rad51 recombinase, which is essential for both mitotic and meiotic recombination. since its discovery, it has been sp ... | 2010 | 20015079 |
| gga autoinhibition revisited. | the cytosolic adaptors gga1-3 mediate sorting of transmembrane proteins displaying a c-terminal acidic dileucine motif (dxxll) in their cytosolic domain. gga1 and gga3 contain similar but intrinsic motifs that are believed to serve as autoinhibitory sites activated by the phosphorylation of a serine positioned three residues upstream of the dxxll motif. in the present study, we have subjected the widely acknowledged concept of gga1 autoinhibition to a thorough structural and functional examinati ... | 2010 | 20015111 |
| human synthetic lethal inference as potential anti-cancer target gene detection. | two genes are called synthetic lethal (sl) if mutation of either alone is not lethal, but mutation of both leads to death or a significant decrease in organism's fitness. the detection of sl gene pairs constitutes a promising alternative for anti-cancer therapy. as cancer cells exhibit a large number of mutations, the identification of these mutated genes' sl partners may provide specific anti-cancer drug candidates, with minor perturbations to the healthy cells. since existent sl data is mainly ... | 2009 | 20015360 |
| interaction between sdo1p and btn1p in the saccharomyces cerevisiae model for batten disease. | juvenile batten disease is an autosomal recessive pediatric neurodegenerative disorder caused by mutations in the cln3 gene. the cln3 protein primarily resides in the lysosomal membrane, but its function is unknown. we demonstrate that cln3 interacts with sbds, the protein mutated in shwachman-bodian-diamond syndrome patients. we demonstrate that this protein-protein interaction is conserved between btn1p and sdo1p, the respective yeast saccharomyces cerevisiae orthologs of cln3 and sbds. it was ... | 2010 | 20015955 |
| nucleosome positioning by genomic excluding-energy barriers. | recent genome-wide nucleosome mappings along with bioinformatics studies have confirmed that the dna sequence plays a more important role in the collective organization of nucleosomes in vivo than previously thought. yet in living cells, this organization also results from the action of various external factors like dna-binding proteins and chromatin remodelers. to decipher the code for intrinsic chromatin organization, there is thus a need for in vitro experiments to bridge the gap between comp ... | 2009 | 20018700 |
| development of a recombinant yeast assay to detect ah-receptor ligands. | endocrine systems of humans and animals are disturbed by dioxin-like compounds, which are ligands of the aryl hydrocarbon receptor (ahr). it is important to determine the accumulation of dioxin-like compounds in the environment for maintenance of human health. in this study, we developed a new method for screening ligands of the ahr using a yeast hybrid system. reporter genes constructed by the insertion of dioxin response elements were integrated into his and lacz yeast genomes. then yeast was ... | 2006 | 20021027 |
| the life span-prolonging effect of sirtuin-1 is mediated by autophagy. | the life span of various model organisms can be extended by caloric restriction as well as by autophagy-inducing pharmacological agents. life span-prolonging effects have also been observed in yeast cells, nematodes and flies upon the overexpression of the deacetylase sirtuin-1. intrigued by these observations and by the established link between caloric restriction and sirtuin-1 activation, we decided to investigate the putative implication of sirtuin-1 in the response of human cancer cells and ... | 2010 | 20023410 |
| modulation of plagl2 transactivation by positive cofactor 2 (pc2), a component of the arc/mediator complex. | the pleomorphic adenoma gene (plag) family of transcription factors regulates a wide range of physiological processes, including cell proliferation, tissue-specific gene regulation, and embryonic development, although little is known regarding the mechanisms that regulate plag protein activity. in this study, a yeast two-hybrid screen identified pc2, a component of the mediator complex, as a plagl2-binding protein. we show that pc2 cooperates with plagl2 and pu.1 to enhance the activity of a kno ... | 2010 | 20025940 |
| cryo-electron tomography of microtubule-kinesin motor complexes. | microtubules complexed with molecular motors of the kinesin family or non-motor microtubule associated proteins (maps) such as tau or eb1 have been the subject of cryo-electron microcopy based 3-d studies for several years. most of these studies that targeted complexes with intact microtubules have been carried out by helical 3-d reconstruction, while few were analyzed by single particle approaches or from 2-d crystalline arrays. helical reconstruction of microtubule-map or motor complexes has b ... | 2010 | 20025975 |
| ptdins4p recognition by vps74/golph3 links ptdins 4-kinase signaling to retrograde golgi trafficking. | targeting and retention of resident integral membrane proteins of the golgi apparatus underly the function of the golgi in glycoprotein and glycolipid processing and sorting. in yeast, steady-state golgi localization of multiple mannosyltransferases requires recognition of their cytosolic domains by the peripheral golgi membrane protein vps74, an orthologue of human golph3/gpp34/gmx33/midas (mitochondrial dna absence sensitive factor). we show that targeting of vps74 and golph3 to the golgi appa ... | 2009 | 20026658 |
| fatty acid synthase impacts the pathobiology of candida parapsilosis in vitro and during mammalian infection. | cytosolic fungal fatty acid synthase is composed of two subunits alpha and beta, which are encoded by fas1 and fas2 genes. in this study, the fas2 genes of the human pathogen candida parapsilosis were deleted using a modified sat1 flipper technique. cpfas2 was essential in media lacking exogenous fatty acids and the growth of fas2 disruptants (fas2 ko) was regulated by the supplementation of different long chain fatty acids, such as myristic acid (14ratio0), palmitic acid (16ratio0), and tween 8 ... | 2009 | 20027295 |
| characterization of a secreted aspartyl protease of the fungal pathogen paracoccidioides brasiliensis. | paracoccidioides brasiliensis is a thermally dimorphic fungus that causes paracoccidioidomycosis, a human systemic disease prevalent in latin america. proteases have been described as playing an important role in the host invasion process in many pathogenic microorganisms. here we describe the identification and characterization of a secreted aspartyl protease (pbsap), isolated from a cdna library constructed with rnas of mycelia transitioning to yeast cells. recombinant pbsap was produced in es ... | 2009 | 20028235 |
| interaction between hhex and sox13 modulates wnt/tcf activity. | fine-tuning of the wnt/tcf pathway is crucial for multiple embryological processes, including liver development. here we describe how the interaction between hhex (hematopoietically expressed homeobox) and sox13 (sry-related high mobility group box transcription factor 13), modulates wnt/tcf pathway activity. hhex is a homeodomain factor expressed in multiple endoderm-derived tissues, like the liver, where it is essential for proper development. the pleiotropic expression of hhex during embryoni ... | 2010 | 20028982 |
| [screening of proteins binding to fxr1p using yeast two-hybrid technique]. | to screen the proteins interacting with fxr1p for functional investigation of fxr1p. | 2009 | 20034884 |
| keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering lrp16 in cytoplasm. | oncogenesis in breast cancer is often associated with excess estrogen receptor alpha(eralpha) activation and overexpression of its coactivators. lrp16 is both an eralpha target gene and an eralpha coactivator, and plays a crucial role in eralpha activation and proliferation of mcf-7 breast cancer cells. however, the regulation of the functional availability of this coactivator protein is not yet clear. | 2009 | 20035625 |