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functional residues on the surface of the n-terminal domain of yeast pms1.saccharomyces cerevisiae mutlalpha is a heterodimer of mlh1 and pms1 that participates in dna mismatch repair (mmr). both proteins have weakly conserved c-terminal regions (ctds), with the ctd of pms1 harboring an essential endonuclease activity. these proteins also have conserved n-terminal domains (ntds) that bind and hydrolyze atp and bind to dna. to better understand pms1 functions and potential interactions with dna and/or other proteins, we solved the 2.5a crystal structure of yeast pms1 ( ...201020138591
structural and functional characterization of recombinant human serum transferrin secreted from pichia pastoris.serum transferrin is an iron-binding glycoprotein with a bilobal structure. it binds iron ions in the blood serum and delivers them into target cells via transferrin receptor. we identified structural and functional characteristics of recombinant human transferrin which is produced in the yeast pichia pastoris. using the signal sequence of the alpha factor of the yeast saccharomyces cerevisiae, high-level secretion was obtained, up to 30 mg/l of culture medium. correct processing at designed sit ...201020139607
an extensive circuitry for cell wall regulation in candida albicans.protein kinases play key roles in signaling and response to changes in the external environment. the ability of candida albicans to quickly sense and respond to changes in its environment is key to its survival in the human host. our guiding hypothesis was that creating and screening a set of protein kinase mutant strains would reveal signaling pathways that mediate stress response in c. albicans. a library of protein kinase mutant strains was created and screened for sensitivity to a variety of ...201020140194
targeted metabolic labeling of yeast n-glycans with unnatural sugars.metabolic labeling of glycans with synthetic sugar analogs has emerged as an attractive means for introducing nonnatural chemical functionality into glycoproteins. however, the complexities of glycan biosynthesis prevent the installation of nonnatural moieties at defined, predictable locations within glycoproteins at high levels of incorporation. here, we demonstrate that the conserved n-acetyglucosamine (glcnac) residues within chitobiose cores of n-glycans in the model organism saccharomyces c ...201020142501
sars coronavirus protein 7a interacts with human ap4a-hydrolase.the sars coronavirus (sars-cov) open reading frame 7a (orf 7a) encodes a 122 amino acid accessory protein. it has no significant sequence homology with any other known proteins. the 7a protein is present in the virus particle and has been shown to interact with several host proteins; thereby implicating it as being involved in several pathogenic processes including apoptosis, inhibition of cellular protein synthesis, and activation of p38 mitogen activated protein kinase. in this study we presen ...201020144233
changes in the proteome of candida albicans in response to azole, polyene, and echinocandin antifungal agents.the yeast candida albicans is an opportunistic human fungal pathogen and the cause of superficial and systemic infections in immunocompromised patients. the classes of antifungal agents most commonly used to treat candida infections are the azoles, polyenes, and echinocandins. in the present study, we identified changes in c. albicans protein abundance using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectroscopy followi ...201020145080
the role of paaac1 encoding a mitochondrial adp/atp carrier in the biosynthesis of extracellular glycolipids, mannosylerythritol lipids, in the basidiomycetous yeast pseudozyma antarctica.pseudozyma antarctica produces large amounts of the glycolipid biosurfactants known as mannosylerythritol lipids (mel), which show not only excellent surface-active properties but also versatile biochemical actions. a gene homologous with a mitochondrial adp/atp carrier was dominantly expressed in p. antarctica under mel-producing conditions on the basis of previous gene expression analysis. the gene encoding the mitochondrial adp/atp carrier of p. antarctica (paaac1) contained a putative open r ...201020146402
assigning roles to dna regulatory motifs using comparative genomics.motivation: transcription factors (tfs) are crucial during the lifetime of the cell. their functional roles are defined by the genes they regulate. uncovering these roles not only sheds light on the tf at hand but puts it into the context of the complete regulatory network. results: here, we present an alignment- and threshold-free comparative genomics approach for assigning functional roles to dna regulatory motifs. we incorporate our approach into the gomo algorithm, a computational tool for d ...201020147307
transgenic mice expressing mutant forms vcp/p97 recapitulate the full spectrum of ibmpfd including degeneration in muscle, brain and bone.inclusion body myopathy associated with paget's disease of bone and frontotemporal dementia (ibmpfd) is a dominantly inherited degenerative disorder caused by mutations in the valosin-containing protein (vcp) gene. vcp (p97 in mouse, ter94 in drosophila melanogaster and cdc48 in saccharomyces cerevisiae) is a highly conserved aaa(+)-atpase that regulates a wide array of cellular processes. the mechanism of ibmpfd pathogenesis is unknown. towards elucidating the pathogenic mechanism we have devel ...201020147319
atr contributes to telomere maintenance in human cells.telomere maintenance is essential to preserve genomic stability and involves several telomere-specific proteins as well as dna replication and repair proteins. the kinase atr, which has a crucial function in maintaining genome integrity from yeast to human, has been shown to be involved in telomere maintenance in several eukaryotic organisms, including yeast, arabidopsis and drosophila. however, its role in telomere maintenance in mammals remains poorly explored. here, we report by using telomer ...201020147462
detection of the atpase activity of the molecular chaperones hsp90 and hsp72 using the transcreenertm adp assay kit.the molecular chaperone heat shock protein 90 (hsp90) is required for the correct folding and stability of a number of client proteins that are important for the growth and maintenance of cancer cells. heat shock protein 72 (hsp72), a co-chaperone of hsp90, is also emerging as an attractive cancer drug target. both proteins bind and hydrolyze adenosine triphosphate (atp), and atpase activity is essential for their function. inhibition of hsp90 atpase activity leads to the degradation of client p ...201020147598
integrated single-cell analysis shows pichia pastoris secretes protein stochastically.the production of heterologous proteins by secretion from cellular hosts is an important determinant for the cost of biotherapeutics. a single-cell analytical method called microengraving was used to examine the heterogeneity in secretion by the methylotrophic yeast pichia pastoris. we show that constitutive secretion of a human fc fragment by p. pastoris is not cell-cycle dependent, but rather fluctuates between states of high and low productivity in a stochastic manner.201020148400
introducing antigen-binding sites in structural loops of immunoglobulin constant domains: fc fragments with engineered her2/neu-binding sites and antibody properties.yeast surface display libraries of human igg1 fc regions were prepared in which loop sequences at the c-terminal tip of the ch3 domain were randomized. a high percentage of these library members bound to soluble cd64 and protein a indicating that the randomization step did not grossly interfere with the overall structure of the displayed fc. sorting these libraries by facs for binders against her2/neu yielded antigen-specific fc binders (fcab; fc antigen binding) of which one was affinity mature ...201020150180
pseudomonas aeruginosa secreted factors impair biofilm development in candida albicans.signal-mediated interactions between the human opportunistic pathogens pseudomonas aeruginosa and candida albicans affect virulence traits in both organisms. phenotypic studies revealed that bacterial supernatant from four p. aeruginosa strains strongly reduced the ability of c. albicans to form biofilms on silicone. this was largely a consequence of inhibition of biofilm maturation, a phenomenon also observed with supernatant prepared from non-clinical bacterial species. the effects of supernat ...201020150241
production of heterologous proteins in plants: strategies for optimal expression.plants are a promising expression system for the production of heterologous proteins, especially therapeutic proteins. currently the majority of therapeutic proteins are produced in mammalian cell lines or bacteria. in a few cases insects, yeast and fungi have been developed for production of human proteins. however, these expression systems have limitations in terms of suitability, cost, scalability, purification and post-translational modifications. therefore, alternative expression systems ar ...201020152894
mechanisms of recombination between diverged sequences in wild-type and blm-deficient mouse and human cells.double-strand breaks (dsbs) are particularly deleterious dna lesions for which cells have developed multiple mechanisms of repair. one major mechanism of dsb repair in mammalian cells is homologous recombination (hr), whereby a homologous donor sequence is used as a template for repair. for this reason, hr repair of dsbs is also being exploited for gene modification in possible therapeutic approaches. hr is sensitive to sequence divergence, such that the cell has developed ways to suppress recom ...201020154148
detection of novel functional selectivity at m3 muscarinic acetylcholine receptors using a saccharomyces cerevisiae platform."functional selectivity", although new to many chemists and biologists only a few years ago, has now become a dominant theme in drug discovery. this concept posits that different ligands engender unique receptor conformations such that only a subset of signaling pathways linked to a given receptor are recruited. however, successful exploitation of the phenomenon to achieve pathway-based selectivity requires the ability to routinely detect it when assessing ligand behavior. we have utilized diffe ...201020155933
hrmas 1h nmr conformational study of the resin-bound amyloid-forming peptide gnnqqny from the yeast prion sup35.the conversion of soluble proteins to insoluble amyloid fibrils is associated with numerous human diseases. the peptide gnnqqny is a short segment of the yeast prion protein sup35 that previously has been found to form amyloid fibrils in a similar manner to the protein itself. the approach taken in this work was to attach this peptide sequence to an insoluble polymer matrix through solid phase peptide synthesis and give it the internal freedom to fold into its local conformation in an organic so ...201020155963
affinity maturation of human botulinum neurotoxin antibodies by light chain shuffling via yeast mating.botulism is caused by the botulinum neurotoxins (bonts), the most poisonous substance known. because of the high potency of bont, development of diagnostic and therapeutic antibodies for botulism requires antibodies of very high affinity. here we report the use of yeast mating to affinity mature bont antibodies by light chain shuffling. a library of immunoglobulin light chains was generated in a yeast vector where the light chain is secreted. the heavy chain variable region and the first domain ...201020156888
comparison of initial hydrolysis of the three dimethyl phthalate esters (dmpes) by a basidiomycetous yeast, trichosporon dmi-5-1, from coastal sediment.purpose: dimethyl phthalate esters (dmpes) are a group of plasticizers commonly detected in the environment with potential adverse human health impact. the degradation of dmpes by fungal systems has been studied to a limited extent, particularly by yeasts. in this study, a basidiomycetous yeast trichosporon dmi-5-1 capable of degrading dmpes was obtained and the degradation pathways were investigated. results: a dmpe-degrading yeast was isolated from costal sediment by enrichment culture techniq ...201121626443
abnormal mitoferrin-1 expression in patients with erythropoietic protoporphyria.objective: most patients with erythropoietic protoporphyria have deficient ferrochelatase (fech) activity due to changes in fech dna. we evaluated seven patients with erythropoietic protoporphyria phenotype in whom abnormalities of fech dna were not found by conventional analysis. the major focus was mitoferrin-1 (mfrn1), the mitochondrial transporter of fe used for heme formation by fech and for 2fe2s cluster synthesis, which is critical to fech activity/stability. materials and methods: four p ...201121627978
yapsin 1 immunoreactivity in alpha cells of human pancreatic islets: implications for the processing of human proglucagon by mammalian aspartic proteases.yapsin 1 is an aspartic protease from saccharomyces cerevisiae and belongs to a class of aspartic proteases that demonstrate specificity for basic amino acids. it is capable of processing prohormone substrates at specific basic residue cleavage sites, similar to that of the prohormone convertases, to generate bioactive peptide hormones. an antibody raised against yapsin 1 was previously shown to immuno-stain endocrine cells of rat pituitary and brain as well as lysates from bovine pituitary secr ...201121632904
over-expression of human calnexin in yeast improves measles surface glycoprotein solubility.the limitations of high-level expression of virus surface proteins in yeast are not well understood. the inefficiency of yeast to produce active human virus surface glycoproteins, as well as other mammalian glycoproteins, is usually explained by the inefficient folding of the glycoprotein into its characteristic and functional three-dimensional structure from a random coil. the endoplasmic reticulum is a highly versatile protein factory that is equipped with chaperones and folding enzymes essent ...201121635688
binding site prediction for protein-protein interactions and novel motif discovery using re-occurring polypeptide sequences.abstract: background: while there are many methods for predicting protein-protein interaction, very few can determine the specific site of interaction on each protein. characterization of the specific sequence regions mediating interaction (binding sites) is crucial for an understanding of cellular pathways. experimental methods often report false binding sites due to experimental limitations, while computational methods tend to require data which is not available at the proteome-scale. here we ...201121635751
structure of the guanylyltransferase domain of human mrna capping enzyme.the enzyme guanylyltransferase (gtase) plays a central role in the three-step catalytic process of adding an (m7)gpppn cap cotranscriptionally to nascent mrna (pre-mrnas). the 5'-mrna capping process is functionally and evolutionarily conserved from unicellular organisms to human. however, the gtases from viruses and yeast have low amino acid sequence identity (∼25%) with gtases from mammals that, in contrast, are highly conserved (∼98%). we have defined by limited proteolysis of human capping e ...201121636784
field testing of different chemical combinations as odour baits for trapping wild mosquitoes in the gambia.odour baited traps have potential use in population surveillance of insect vectors of disease, and in some cases for vector population reduction. established attractants for human host-seeking mosquitoes include a combination of co(2) with l-lactic acid and ammonia, on top of which additional candidate compounds are being tested. in this field study in rural gambia, using latin square experiments with thorough randomization and replication, we tested nine different leading candidate combinations ...201121637337
an acidic loop and cognate phosphorylation sites define a molecular switch that modulates ubiquitin charging activity in cdc34-like enzymes.e2 ubiquitin-conjugating enzymes are crucial mediators of protein ubiquitination, which strongly influence the ultimate fate of the target substrates. recently, it has been shown that the activity of several enzymes of the ubiquitination pathway is finely tuned by phosphorylation, an ubiquitous mechanism for cellular regulation, which modulates protein conformation. in this contribution, we provide the first rationale, at the molecular level, of the regulatory mechanism mediated by casein kinase ...201121637798
gene network landscape of the ciliate tetrahymena thermophila.genome-wide expression data of gene microarrays can be used to infer gene networks. at a cellular level, a gene network provides a picture of the modules in which genes are densely connected, and of the hub genes, which are highly connected with other genes. a gene network is useful to identify the genes involved in the same pathway, in a protein complex or that are co-regulated. in this study, we used different methods to find gene networks in the ciliate tetrahymena thermophila, and describe s ...201121637855
involvement of a toxoplasma gondii chromatin remodeling complex ortholog in developmental regulation.the asexual cycle of the parasite toxoplasma gondii has two developmental stages: a rapidly replicating form called a tachyzoite and a slow growing cyst form called a bradyzoite. while the importance of atp-independent histone modifications for gene regulation in t. gondii have been demonstrated, atp-dependent chromatin remodeling pathways have not been examined. in this study we characterized c9, an insertional mutant showing reduced expression of bradyzoite differentiation marker bag1, in cult ...201121655329
calcium signaling components in the human pathogen: cryptococcus neoformans.calcium signaling through calmodulin and the phosphatase calcineurin are required for key events of the biology of the human pathogen cryptococcus neoformans, including mating, morphogenesis, growth at 37°c and virulence. in a recent work we described the functional characterization of a new component of this calcium signaling network: the vacuolar calcium exchanger vcx1. this transporter is involved in calcium tolerance and virulence in c. neoformans. two other uncharacterized calcium transport ...201121655435
the over-expression of the ╬▓2 catalytic subunit of the proteasome decreases homologous recombination and impairs dna double-strand break repair in human cells.by a human cdna library screening, we have previously identified two sequences coding two different catalytic subunits of the proteasome which increase homologous recombination (hr) when overexpressed in the yeast saccharomyces cerevisiae. here, we investigated the effect of proteasome on spontaneous hr and dna repair in human cells. to determine if the proteasome has a role in the occurrence of spontaneous hr in human cells, we overexpressed the ╬▓2 subunit of the proteasome in hela cells and d ...201121660142
in vivo interaction of the hepatitis delta virus small antigen with the elav-like protein hur.the small and large delta antigens (s-hdag and l-hdag, respectively) represent two forms of the only protein encoded by the hepatitis delta virus (hdv) rna genome. consequently, hdv relies, at a large extent, on the host cell machinery for replication and transcription. until now, only a limited number of cellular proteins were identified as s-hdag or l-hdag partners being involved in the modulation of the virus life cycle. in an attempt to identify cellular s-hdag-binding proteins we made use o ...201121660185
model organisms reveal insight into human neurodegenerative disease: ataxin-2 intermediate-length polyglutamine expansions are a risk factor for als.model organisms include yeast saccromyces cerevisae and fly drosophila melanogaster. these systems have powerful genetic approaches, as well as highly conserved pathways, both for normal function and disease. here, we review and highlight how we applied these systems to provide mechanistic insight into the toxicity of tdp-43. tdp-43 accumulates in pathological aggregates in als and about half of ftd. yeast and fly studies revealed an interaction with the counterparts of human ataxin-2, a gene wh ...201121660502
do cancer proteins really interact strongly in the human protein-protein interaction network?protein-protein interaction (ppi) network analysis has been widely applied in the investigation of the mechanisms of diseases, especially cancer. recent studies revealed that cancer proteins tend to interact more strongly than other categories of proteins, even essential proteins, in the human interactome. however, it remains unclear whether this observation was introduced by the bias towards more cancer studies in humans. here, we examined this important issue by uniquely comparing network char ...201121666777
osbp-related protein 7 interacts with gate-16 and negatively regulates gs28 protein stability.orp7 is a member of oxysterol-binding protein (osbp) family, the function of which has remained obscure. in this study, we identified by yeast two-hybrid screening an interaction partner of orp7, gate-16, which (i) regulates golgi snare of 28kda (gs28) function and stability, and (ii) plays a role in autophagosome biogenesis. the interaction was confirmed by bimolecular fluorescence complementation (bifc) assay in living cells. the interacting regions were delineated within aa 1-142 of orp7 and ...201121669198
insights into the structure of the ccr4-not complex by electron microscopy.the ccr4-not complex is a deadenylation complex, which plays a major role for mrna stability. the complex is conserved from yeast to human and consists of nine proteins not1-not5, ccr4, caf1, caf40 and caf130. we have successfully isolated the complex using a protein a tag on not1, followed by cross-linking on a glycerol gradient. all components of the complex were identified by mass spectrometry. electron microscopy of negatively stained particles followed by image reconstruction revealed an l- ...201121669201
substitution patterns are gc-biased in divergent sequences across the metazoans.the fastest-evolving regions in the human and chimpanzee genomes show a remarkable excess of weak (a,t) to strong (g,c) nucleotide substitutions since divergence from their common ancestor. we investigated the phylogenetic extent and possible causes of this weak to strong (w ôåæ s) bias in divergent sequences (bds) using recently sequenced genomes and recombination maps from eight trios of eukaryotic species. to quantify evidence for bds, we inferred substitution histories using an efficient max ...201121670083
fission yeast homologs of human xpc and csb, rhp41 and rhp26, are involved in transcription-coupled repair of methyl methanesulfonate-induced dna damage.methyl methanesulfonate (mms) methylates nitrogen atoms in purines, and predominantly produces 7-methylguanine and 3-methyladenine (3-mea). previously, we showed that base excision repair (ber) and nucleotide excision repair (ner) synergistically function to repair mms-induced dna damage in the fission yeast schizosaccharomyces pombe. here, we studied the roles of ner components in repair of 3-mea and ber intermediates such as the ap site and single strand breaks. mutants of rhp41 (xpc homolog) ...201121670547
p53 transactivation and the impact of mutations, cofactors and small molecules using a simplified yeast-based screening system.the p53 tumor suppressor, which is altered in most cancers, is a sequence-specific transcription factor that is able to modulate the expression of many target genes and influence a variety of cellular pathways. inactivation of the p53 pathway in cancer frequently occurs through the expression of mutant p53 protein. in tumors that retain wild type p53, the pathway can be altered by upstream modulators, particularly the p53 negative regulators mdm2 and mdm4.201121674059
plant ornithine decarboxylase is not post-transcriptionally feedback regulated by polyamines but can interact with a cytosolic ribosomal protein s15 polypeptide.the formation of putrescine by ornithine decarboxylase (odc) is a key regulatory step in polyamine biosynthesis in metazoa and fungi. excess polyamines post-transcriptionally induce the synthesis of a unique non-competitive protein inhibitor of odc, termed antizyme. binding of antizyme to an odc monomer subunit results in enzymatic inhibition, rapid ubiquitin-independent degradation of odc by the 26s proteasome and recycling of antizyme. plants possess an additional route for synthesizing putres ...201121814791
detection of antibodies against paracoccidioides brasiliensis melanin: in vitro and in vivo studies during infection.several cell-wall constituents, including melanins or melanin-like compounds, have been implicated in the pathogenesis of a wide variety of microbial diseases caused by diverse species of pathogenic bacteria, fungi and helminthes. among these microorganisms, the dimorphic fungal pathogen paracoccidioides brasiliensis produces melanin in its conidial and yeast forms. in the present study, melanin particles from p. brasiliensis were injected into balb/c mice in order to produce monoclonal antibodi ...201121813659
human cabin1 is a functional member of the human hira/ubn1/asf1a histone h3.3 chaperone complex.the mammalian hira/ubn1/asf1a complex is a histone chaperone complex that is conserved from yeast to humans. this complex preferentially deposits the histone variant h3.3 into chromatin in a dna replication-independent manner, and is implicated in diverse chromatin regulatory events from gene activation to heterochromatinization. in yeast, the orthologous complex consists of three hir proteins (hir1p, hir2p and hir3p), hpc2p and asf1p. yeast hir3p has weak homology to, cabin1, a fourth member of ...201121807893
phospholipid scramblase 1 mediates hepatitis c virus entry into host cells.hepatitis c virus (hcv) infects human hepatocytes through several host factors. however, other prerequisite factors for viral entry remain to be identified. using a yeast two-hybrid screen, we found that human phospholipid scramblase 1 interacts with hcv envelope proteins e1 and e2. these physical interactions were confirmed by co-immunoprecipitation and gst pull-down assays. knocking down the expression of plscr1 inhibited the entry of hcv pseudoparticles. moreover, plscr1 was required for the ...201121806988
the cellular dexd/h-box rna-helicases uap56 and urh49 exhibit a crm1-independent nucleocytoplasmic shuttling activity.cellular dexd/h-box rna-helicases perform essential functions during mrna biogenesis. the closely related human proteins uap56 and urh49 are members of this protein family and play an essential role for cellular mrna export by recruiting the adaptor protein ref to spliced and unspliced mrnas. in order to gain insight into their mode of action, we aimed to characterize these rna-helicases in more detail. here, we demonstrate that uap56 and urh49 exhibit an intrinsic crm1-independent nucleocytopla ...201121799930
ddx60, a dexd/h box helicase, is a novel antiviral factor promoting rig-i-like receptor-mediated signaling.the cytoplasmic viral rna sensors rig-i and mda5 are important for the production of type i interferon and other inflammatory cytokines. ddx60 is an uncharacterized dexd/h box rna helicase similar to s. cerevisiae ski2, a cofactor of rna exosome, a protein complex required for the integrity of cytoplasmic rna. the expression of ddx60 increases after viral infection, and the protein localizes at cytoplasmic region. after viral infection, the ddx60 protein binds to endogenous rig-i protein. the pr ...201121791617
papd5, a noncanonical poly(a) polymerase with an unusual rna-binding motif.papd5 is one of the seven members of the family of noncanonical poly(a) polymerases in human cells. papd5 was shown to polyadenylate aberrant pre-ribosomal rnas in vivo, similar to degradation-mediating polyadenylation by the noncanonical poly(a) polymerase trf4p in yeast. papd5 has been reported to be also involved in the uridylation-dependent degradation of histone mrnas. to test whether papd5 indeed catalyzes adenylation as well as uridylation of rna substrates, we analyzed the in vitro prope ...201121788334
identification of brain-specific angiogenesis inhibitor 2 as an interaction partner of glutaminase interacting protein.the vast majority of physiological processes in living cells are mediated by protein-protein interactions often specified by particular protein sequence motifs. pdz domains, composed of 80-100 amino acid residues, are an important class of interaction motif. among the pdz-containing proteins, glutaminase interacting protein (gip), also known as tax interacting protein tip-1, is unique in being composed almost exclusively of a single pdz domain. gip has important roles in cellular signaling, prot ...201121787750
turnover of atp synthase subunits in f(1)-depleted hela and yeast cells.mitochondrial translation of the saccharomyces cerevisiae atp6p subunit of f(1)-f(0) atp synthase is regulated by the f(1) atpase. here we show normal expression of atp6p in hela cells depleted of the f(1) ß subunit. instead of being translationally down-regulated, hela cells lacking f(1) degrade atp6p, thereby preventing proton leakage across the inner membrane. mammalian mitochondria also differ in the way they minimize the harmful effect of unassembled f(1) a subunit. while yeast mutants lack ...201121784071
[activity of macrolides and fluoroquinolones against intracellular legionella pneumophila].to evaluate the activity of macrolides and fluoroquinolones against legionella pneumophila by intracellular susceptibility testing.201121781510
identification of an evolutionary conserved family of inorganic polyphosphate endopolyphosphatases.inorganic polyphosphate (poly-p) consists of just a chain of phosphate groups linked by high-energy bonds. it is found in every organism and is implicated in a wide variety of cellular processes, e.g. phosphate storage, blood coagulation, pathogenicity. its metabolism has been studied mainly in bacteria, while remaining largely uncharacterized in eukaryotes. it has recently been suggested that poly-p metabolism is connected to that of highly phosphorylated inositol species (inositol pyrophosphat ...201121775424
[the effects of tata-box in cyc1 promoter on the reporter gene regulated by ere in the recombinant yeast cell].the complementary oligonucleotides, each with two consensus estrogen response element (ere)-sequences and 5'-hind iii and 3'-sph i sticky ends were artificially synthesized. a solution with both the complementary dna sequences was heated to 95'c and cooled down to room temperature to form double strand dna (dsdna). the set was cloned into the corresponding sites of cyc1 promoter of the pere-cyc-yegfp to yield pere-cycalpha-yegfp vector. the two different reporter vectors, pere-cyc-yegfp and pere ...201121774222
effect of phyllanthus emblica linn. on candida adhesion to oral epithelium and denture acrylic.to investigate the effect of phyllanthus emblica (p. emblica) linn. ethanolic extract on the adhesion of candida albicans (c. albicans) to human buccal epithelial cells (becs) and denture acrylic surfaces.201121771413
type ix collagen interacts with fibronectin providing an important molecular bridge in articular cartilage.type ix collagen is covalently bound to the surface of type ii collagen fibrils within the cartilage ecm. the n-terminal, globular non-collagenous domain (nc4) of the a1(ix) chain protrudes away from the surface of the fibrils into the surrounding matrix and is available for molecular interactions. to define these interactions we used the nc4 domain in a yeast two-hybrid screen of a human chondrocyte cdna library. 73% of the interacting clones encoded fibronectin. the interaction was confirmed u ...201121768108
human fatty acid transport protein 2a/very long chain acyl coa synthetase 1 (fatp2a/acsvl1) has a preference in mediating the channeling of exogenous n-3 fatty acids into phosphatidylinositol.the trafficking of fatty acids across the membrane and into downstream metabolic pathways requires their activation to coa thioesters. members of the fatty acid transport protein/very long chain acyl coa synthetase (fatp/acsvl) family are emerging as key players in the trafficking of exogenous fatty acids into the cell and in intracellular fatty acid homeostasis. we have expressed two naturally occurring splice variants of fatp2 (acsvl1) in yeast and 293t-rex cells and addressed their roles in f ...201121768100
sirtuins, bioageing, and cancer.the sirtuins are a family of orthologues of yeast sir2 found in a wide range of organisms from bacteria to man. they display a high degree of conservation between species, in both sequence and function, indicative of their key biochemical roles. sirtuins are heavily implicated in cell cycle, cell division, transcription regulation, and metabolism, which places the various family members at critical junctures in cellular metabolism. typically, sirtuins have been implicated in the preservation of ...201121766030
characterization of yipf3 and yipf4, cis-golgi localizing yip domain family proteins.the yip1 domain family (yipf) proteins are homologues of yeast yip1p and yif1p, which are proposed to function in er to golgi transport. here, we report the characterization of yipf3 and yipf4, homologues of human yif1p and yip1p, respectively. immunofluorescence and immuno-electron microscopy showed that both yipf3 and yipf4 are clearly concentrated in the cis-golgi. while yipf4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms ...201121757827
cell surface display of functional human mhc class ii proteins: yeast display versus insect cell display.reliable and robust systems for engineering functional major histocompatibility complex class ii (mhcii) proteins have proved elusive. availability of such systems would enable the engineering of peptide-mhcii (pmhcii) complexes for therapeutic and diagnostic applications. in this paper, we have developed a system based on insect cell surface display that allows functional expression of heterodimeric dr2 molecules with or without a covalently bound human myelin basic protein (mbp) peptide, which ...201121752831
the inhibitor of growth protein 5 (ing5) depends on inca1 as a co-factor for its antiproliferative effects.the proteins of the inhibitor of growth (ing) family are involved in multiple cellular functions such as cell cycle regulation, apoptosis, and chromatin remodeling. for ing5, its actual role in growth suppression and the necessary partners are not known. in a yeast-two-hybrid approach with human bone marrow derived cdna, we identified ing5 as well as several other proteins as interaction partners of inhibitor of cyclin a1 (inca1) that we previously characterized as a novel interaction partner of ...201121750715
ploidy influences cellular responses to gross chromosomal rearrangements in saccharomyces cerevisiae.abstract:201121711526
overexpression of a homogeneous oligosaccharide with (13)c labeling by genetically engineered yeast strain.this report describes a novel method for overexpression of (13)c-labeled oligosaccharides using genetically engineered saccharomyces cerevisiae cells, in which a homogeneous high-mannose-type oligosaccharide accumulates because of deletions of genes encoding three enzymes involved in the processing pathway of asparagine-linked oligosaccharides in the golgi complex. using uniformly (13)c-labeled glucose as the sole carbon source in the culture medium of these engineered yeast cells, high yields o ...201121698488
candida albicans hgt1p, a multifunctional evasion molecule: complement inhibitor, cr3 analogue, and human immunodeficiency virus-binding molecule.background.ôçâthe complement system is tightly controlled by several regulators. two of these, factor h (fh) and c4b-binding protein (c4bp), can be acquired by pathogens conveying resistance to complement attack. the aim of the study was to characterize the fh binding molecule of candida albicans, a potentially life-threatening yeast. methods.ôçâthe gene coding for this molecule was identified by probing an expression library and homozygous deletion mutants of the respective gene were constructe ...201121844307
bioorthogonal chemical reporters for analyzing protein lipidation and lipid trafficking.protein lipidation and lipid trafficking control many key biological functions in all kingdoms of life. the discovery of diverse lipid species and their covalent attachment to many proteins has revealed a complex and regulated network of membranes and lipidated proteins that are central to fundamental aspects of physiology and human disease. given the complexity of lipid trafficking and the protein targeting mechanisms involved with membrane lipids, precise and sensitive methods are needed to mo ...201121675729
An essential nuclear protein in trypanosomes is a component of mRNA transcription/export pathway.In eukaryotic cells, different RNA species are exported from the nucleus via specialized pathways. The mRNA export machinery is highly integrated with mRNA processing, and includes a different set of nuclear transport adaptors as well as other mRNA binding proteins, RNA helicases, and NPC-associated proteins. The protozoan parasite Trypanosoma cruzi is the causative agent of Chagas disease, a widespread and neglected human disease which is endemic to Latin America. Gene expression in Trypanosoma ...201121687672
A yeast two hybrid screen identifies SPATA4 as a TRAPP interactor.The TRAPP vesicle-tethering complex consists of more than 10 distinct polypeptides and is involved in protein transport. Using the C2 subunit as bait we identified SPATA4, a spermatocyte-specific protein of unknown function, as an interacting partner in a yeast two hybrid screen. Further studies indicate SPATA4 interacts with the C2 portion of the TRAPP complex. SPATA4 fractionates with both cytosolic and nuclear fractions suggesting it may have several distinct functions. SPATA4 is one of only ...201121827752
Dipoid-specific genome stability genes of S. cerevisiae: genomic screen reveals haploidization as an escape from persisting DNA rearrangement stress.Maintaining a stable genome is one of the most important tasks of every living cell and the mechanisms ensuring it are similar in all of them. The events leading to changes in DNA sequence (mutations) in diploid cells occur one to two orders of magnitude more frequently than in haploid cells. The majority of those events lead to loss of heterozygosity at the mutagenesis marker, thus diploid-specific genome stability mechanisms can be anticipated. In a new global screen for spontaneous loss of fu ...201121695049
p53 gene repair with zinc finger nucleases optimised by yeast 1-hybrid and validated by solexa sequencing.the tumor suppressor gene p53 is mutated or deleted in over 50% of human tumors. as functional p53 plays a pivotal role in protecting against cancer development, several strategies for restoring wild-type (wt) p53 function have been investigated. in this study, we applied an approach using gene repair with zinc finger nucleases (zfns). we adapted a commercially-available yeast one-hybrid (y1h) selection kit to allow rapid building and optimization of 4-finger constructs from randomized pcr libra ...201121695267
species of candida isolated from anatomically distinct sites in military personnel in cuiabá, mato grosso, brazil.some fungi are natural inhabitants of the human body but may result in disease when conditions are conducive to their development. yeast infections are common and often occur in the skin and mucous membranes; however emerging species have changed this epidemiological profile. the ability to colonize different anatomical sites has been associated with the pathogenicity of candida when environmental conditions are particularly favorable. in the case of hot, humid climates, the attrition suffered b ...201121987132
ATM controls meiotic double-strand-break formation.In many organisms, developmentally programmed double-strand breaks (DSBs) formed by the SPO11 transesterase initiate meiotic recombination, which promotes pairing and segregation of homologous chromosomes. Because every chromosome must receive a minimum number of DSBs, attention has focused on factors that support DSB formation. However, improperly repaired DSBs can cause meiotic arrest or mutation; thus, having too many DSBs is probably as deleterious as having too few. Only a small fraction of ...201122002603
A role for SUMO in nucleotide excision repair.The two Siz/PIAS SUMO E3 ligases Siz1 and Siz2 are responsible for the vast majority of sumoylation in Saccharomyces cerevisiae. We found that siz1? siz2? mutants are sensitive to ultra-violet (UV) light. Epistasis analysis showed that the SIZ genes act in the nucleotide excision repair (NER) pathway, and suggested that they participate both in global genome repair (GGR) and in the Rpb9-dependent subpathway of transcription-coupled repair (TCR), but have minimal role in Rad26-dependent TCR. Quan ...201121968059
The plasma membrane-associated GTPase Rin interacts with the dopamine transporter and is required for protein kinase C-regulated dopamine transporter trafficking.Dopaminergic signaling and plasticity are essential to numerous CNS functions and pathologies, including movement, cognition, and addiction. The amphetamine- and cocaine-sensitive dopamine (DA) transporter (DAT) tightly controls extracellular DA concentrations and half-life. DAT function and surface expression are not static but are dynamically modulated by membrane trafficking. We recently demonstrated that the DAT C terminus encodes a PKC-sensitive internalization signal that also suppresses b ...201121957239
MIF from mussel: Coding sequence, phylogeny, polymorphism, 3D model and regulation of expression.Three macrophage migration inhibitory factor (MIF)-related sequences were identified from a Mytilus galloprovincialis EST library. The consensus sequence included a 5'-UTR of 32 nucleotides, the complete ORF of 345 nucleotides, and a 3'-UTR of 349 nucleotides. As for other MIFs, M. galloprovincialis ORF does not include any signal or C-terminus extensions. The translated sequence of 115 amino acids possesses a molecular mass of 12,681.4, a pI of 6.27 and a stability index of 21.48. Its 3D struct ...201122085783
Antagonists of anaphase-promoting complex (APC)-2-cell cycle and apoptosis regulatory protein (CARP)-1 interaction are novel regulators of cell growth and apoptosis.CARP-1/CCAR1, a perinuclear phosphoprotein, is a regulator of cell growth and apoptosis signaling. Although CARP-1 is a regulator of chemotherapy-dependent apoptosis, it is also a part of the NF-?B proteome and a co-activator of steroid/thyroid nuclear receptors as well as ß-catenin signaling. Our yeast two-hybrid screen revealed CARP-1 binding with the anaphase-promoting complex/cyclosome E3 ubiquitin ligase component APC-2 protein. CARP-1 also binds with anaphase-promoting complex/cyclosome co ...201121903591
Biochemical characterization of Warsaw Breakage Syndrome helicase.Mutations in the human ChlR1 gene are associated with a unique genetic disorder known as Warsaw Breakage Syndrome characterized by cellular defects in sister chromatid cohesion and hypersensitivity to agents that induce replication stress. A role of ChlR1 helicase in sister chromatid cohesion was first evidenced by studies of the yeast homolog Chl1p; however, its cellular functions in DNA metabolism are not well understood. We have carefully examined the DNA substrate specificity of purified r ...201122102414
toward an understanding of the protein interaction network of the human liver.proteome-scale protein interaction maps are available for many organisms, ranging from bacteria, yeast, worms and flies to humans. these maps provide substantial new insights into systems biology, disease research and drug discovery. however, only a small fraction of the total number of human protein-protein interactions has been identified. in this study, we map the interactions of an unbiased selection of 5026 human liver expression proteins by yeast two-hybrid technology and establish a human ...201121988832
air1 zinc knuckles 4 and 5 and a conserved iwrxy motif are critical for the function and integrity of the trf4/5-air1/2-mtr4 polyadenylation (tramp) rna quality control complex.in saccharomyces cerevisiae, non-coding rnas, including cryptic unstable transcripts (cuts), are subject to degradation by the exosome. the trf4/5-air1/2-mtr4 polyadenylation (tramp) complex in s. cerevisiae is a nuclear exosome cofactor that recruits the exosome to degrade rnas. trf4/5 are poly(a) polymerases, mtr4 is an rna helicase, and air1/2 are putative rna-binding proteins that contain five cchc zinc knuckles (znks). one central question is how the tramp complex, especially the air1/2 pro ...201121878619
Fulminant Candidemia Diagnosed by Prompt Detection of Pseudohyphae in a Peripheral Blood Smear.A 77-year-old man treated with prednisolone for pemphigus developed severe sepsis by Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. Several antibiotics were administered. A peripheral blood smear showed growth of a large number of yeast extending pseudohyphae which could be seen both inside and outside of leucocytes. Antifungal agents were added immediately; however, he did not recover. Several days later, blood culture showed Candida albicans septicemia. The autopsy rev ...201122173051
plasticity and diversity of trna anticodon determinants of substrate recognition by eukaryotic a37 isopentenyltransferases.the n(6)-(isopentenyl)adenosine (i(6)a) modification of some trnas at position a37 is found in all kingdoms and facilitates codon-specific mrna decoding, but occurs in different subsets of trnas in different species. here we examine yeasts' trna isopentenyltransferases (i.e., dimethylallyltransferase, dmatase, members of the δ(2)-isopentenylpyrophosphate transferase, ippt superfamily) encoded by tit1(+) in schizosaccharomyces pombe and mod5 in saccharomyces cerevisiae, whose homologs are escheri ...201121873461
inhibition of homologous recombination by the pcna-interacting protein pari.inappropriate homologous recombination (hr) causes genomic instability and cancer. in yeast, the uvrd family helicase srs2 is recruited to sites of dna replication by sumo-modified pcna, where it acts to restrict hr by disassembling toxic rad51 nucleofilaments. how human cells control recombination at replication forks is unknown. here, we report that the protein pari, containing a uvrd-like helicase domain, is a pcna-interacting partner required for preservation of genome stability in human and ...201122153967
Structural analysis of the interaction between Hsp90 and the tumor suppressor protein p53.In eukaryotes, the essential dimeric molecular chaperone Hsp90 is required for the activation and maturation of specific substrates such as steroid hormone receptors, tyrosine kinases and transcription factors. Hsp90 is involved in the establishment of cancer and has become an attractive target for drug design. Here we present a structural characterization of the complex between Hsp90 and the tumor suppressor p53, a key mediator of apoptosis whose structural integrity is crucial for cell-cycle c ...201121892170
probing the force generation and stepping behavior of cytoplasmic dynein.cytoplasmic dynein, which is the largest and arguably the most complex cytoskeletal motor protein, plays fundamental roles during cell division, nuclear positioning, and organelle and mrna transport, by generating force and movement toward the minus ends of microtubules. consequently, dynein is central to many physiological processes, and its dysfunction is implicated in human diseases. however, the molecular mechanism by which dynein produces force and movement remains poorly understood. here, ...201121909883
hydroxylation of recombinant human collagen type i alpha 1 in transgenic maize co-expressed with a recombinant human prolyl 4-hydroxylase.abstract:201121702901
opposing effects of glutamine and asparagine govern prion formation by intrinsically disordered proteins.sequences rich in glutamine (q) and asparagine (n) residues often fail to fold at the monomer level. this, coupled to their unusual hydrogen-bonding abilities, provides the driving force to switch between disordered monomers and amyloids. such transitions govern processes as diverse as human protein-folding diseases, bacterial biofilm assembly, and the inheritance of yeast prions (protein-based genetic elements). a systematic survey of prion-forming domains suggested that q and n residues have d ...201121726811
the ubc9 e2 sumo conjugating enzyme binds the pr-set7 histone methyltransferase to facilitate target gene repression.pr-set7/set8/kmt5a is a chromatin-modifying enzyme that specifically monomethylates lysine 20 of histone h4 (h4k20me1). in this study we attempted to identify pr-set7-interacting proteins reasoning that these proteins would provide important insights into the role of pr-set7 in transcriptional regulation. using an unbiased yeast two-hybrid approach, we discovered that pr-set7 interacts with the ubc9 e2 sumo conjugating enzyme. this interaction was confirmed in human cells and we demonstrated tha ...201121829513
sirt6 promotes dna repair under stress by activating parp1.sirtuin 6 (sirt6) is a mammalian homolog of the yeast sir2 deacetylase. mice deficient for sirt6 exhibit genome instability. here, we show that in mammalian cells subjected to oxidative stress sirt6 is recruited to the sites of dna double-strand breaks (dsbs) and stimulates dsb repair, through both nonhomologous end joining and homologous recombination. our results indicate that sirt6 physically associates with poly[adenosine diphosphate (adp)-ribose] polymerase 1 (parp1) and mono-adp-ribosylate ...201121680843
[new nutrient medium for the cultivation and isolation of the plague microbe chds-37 as an element of the mobilization reserve of specialized antiepidemic teams of the russian inspectorate for the protection of consumer rights and human welfare].a new nutrient medium has been designed to culture and isolate the plague microbe chds-37 on the basis of the pancreatic digest of baker's yeast. the results of laboratory tests of the designed medium, by using 10 plague microbe strains and those of approval during the tactical and special training of a specialized antiepidemic team (saet), suggest that the medium has some advantage over reference media and creates prerequisites for being incorporated into the mobilization reserve of a saet.201121735682
isolation of compensatory inhibitor domain mutants to novel activation domain variants using the split-ubiquitin screen.the control of transcription factor function plays an important role in the development of many processes in eukaryotes, such as drug resistance in fungi and human tumours undergoing chemotherapy. detailed molecular mapping of the interactions between transcription factors and their protein partners can give important information about their mechanisms of action and reveal potential therapeutic targets. we devised a genetic screening system for mapping the interaction site between the saccharomy ...201121732556
three "hotspots" important for adenosine a(2b) receptor activation: a mutational analysis of transmembrane domains 4 and 5 and the second extracellular loop.g protein-coupled receptors (gpcrs) are a major drug target and can be activated by a range of stimuli, from photons to proteins. despite the progress made in the last decade in molecular and structural biology, their exact activation mechanism is still unknown. here we describe new insights in specific regions essential in adenosine a(2b) receptor activation (a(2b)r), a typical class a gpcr. we applied unbiased random mutagenesis on the middle part of the human adenosine a(2b)r, consisting of t ...201121818573
tmp21, a novel mhc-i interacting protein, preferentially binds to +æ2-microglobulin-free mhc-i heavy chains.mhc-i molecules play a critical role in immune surveillance against viruses by presenting peptides to cytotoxic t lymphocytes. although the mechanisms by which mhc-i molecules assemble and acquire peptides in the er are well characterized, how mhc-i molecules traffic to the cell surface remains poorly understood. to identify novel proteins that regulate the intracellular transport of mhc-i molecules, mhc-i-interacting proteins were isolated by affinity purification, and their identity was determ ...201121699748
legionella pneumophilarequires polyamines for optimal intracellular growth.the gram-negative intracellular pathogen legionella pneumophilareplicates in a membrane-bound compartment known as the legionella-containing vacuole (lcv), into which it abundantly releases its chaperonin, htpb. to determine whether htpb remains within the lcv or reaches the host cell cytoplasm, we infected u937 human macrophages and cho cells with l. pneumophilaexpressing a translocation reporter consisting of the bordetella pertussisadenylate cyclase fused to htpb. these infections led to incr ...201121742865
candida glabrata pwp7p and aed1p are required for adherence to human endothelial cells.candida glabrata owes its success as a pathogen, in part, to a large repertoire of adhesins present on the cell surface. our current knowledge of c. glabrata adhesins and their role in the interaction between host and pathogen is limited to work with only a single family of epithelial adhesins (epa proteins). here, we report on the identification and characterization of a family of glycosylphosphatidylinositol-anchored cell wall proteins in c. glabrata. these proteins are absent in both saccharo ...201121726406
fly meets yeast: checking the correct orientation of cell division.cell division is generally thought to be a process that produces an exact copy of the mother cell by precisely replicating its genomic dna, doubling organelles, and segregating them into two cells. many cell types from bacteria to human cells divide asymmetrically, however, to generate daughter cells with distinct characteristics. such asymmetric divisions are fundamental to the lifespan of a cell, to embryonic development, and to stem cell homeostasis. asymmetric division requires coordination ...201121705221
targeting the regulation of androgen receptor signaling by the heat shock protein 90 cochaperone fkbp52 in prostate cancer cells.drugs that target novel surfaces on the androgen receptor (ar) and/or novel ar regulatory mechanisms are promising alternatives for the treatment of castrate-resistant prostate cancer. the 52 kda fk506 binding protein (fkbp52) is an important positive regulator of ar in cellular and whole animal models and represents an attractive target for the treatment of prostate cancer. we used a modified receptor-mediated reporter assay in yeast to screen a diversified natural compound library for inhibito ...201121730179
stereoselective biotransformation of timosaponin a-iii by saccharomyces cerevisiae.bioconversion of timosaponin a-iii (ta-iii), one of the major steroidal saponins isolated from the rhizomes of anemarrhenae asphodeloides bunge (liliaceae), was investigated in saccharomyces cerevisiae. five bioconversion products, denoted compounds 2-6, were obtained. biotransformation metabolite 2 was a stereoisomer of taiii with a specific isotype f-ring and beta-ranged ch3-21, which rarely occurs in nature. the structure of 2 was elucidated by extensive spectroscopic analysis (h-h cosy, hsqc ...201121715964
feedback regulation of ran gene expression by ran protein.ran, an abundant gtpase that is highly conserved in eukaryotes from yeast to human, has been implicated in many aspects of nuclear structure and function. recently it is revealed that the ran gtpase can regulate the hemocytic phagocytosis of shrimp by interaction with myosin. however the regulation of ran gene expression remains unknown. in this study, the promoter of shrimp ran gene was identified which contained a typical tata box. the results showed that the shrimp ran protein was bound with ...201121741460
saccharomyces as a vaccine against systemic aspergillosis: 'the friend of man' a friend again?the mortality of clinical aspergillus infections necessitates consideration of the utility of a vaccine. we have found that saccharomyces species can act as a protective vaccine against a lethal systemic aspergillus infection, and describe experiments optimizing a subcutaneous regimen with killed yeast. three injections of 2.5 mg given a week apart, 2 weeks prior to challenge, consistently, significantly, provided survival protection and reduction of infection in organs in survivors. the protect ...201121825307
purification and characterization of a novel aspartic protease from basidiomycetous yeast cryptococcus sp. s-2.an aspartic protease (cap1) was purified from basidiomycetous yeast cryptococcus sp. s-2 (ferm abp-10961) using hitrap deae ff column and hitrap q hp column chromatography with azocasein as a substrate. cap1 has a molecular mass of 34kda on sds-page. it was stable up to 50-¦c with maximum activity at 30-¦c. maximum proteolytic activity was observed at ph 5.0. cap1 was stable in the ph range 3.0-7.0. its enzyme activity was strongly inhibited by pepstatin a, an inhibitor of aspartic proteases, in ...201121824815
mediator and human disease.since the identification of a metazoan counterpart to yeast mediator nearly 15 years ago, a convergent body of biochemical and molecular genetic studies have confirmed their structural and functional relationship as an integrative hub through which regulatory information conveyed by signal activated transcription factors is transduced to rna polymerase ii. nonetheless, metazoan mediator complexes have been shaped during evolution by substantive diversification and expansion in both the number an ...201121840410
kif1a, an axonal transporter of synaptic vesicles, is mutated in hereditary sensory and autonomic neuropathy type 2.hereditary sensory and autonomic neuropathy type ii (hsanii) is a rare autosomal-recessive disorder characterized by peripheral nerve degeneration resulting in a severe distal sensory loss. although mutations in fam134b and the hsn2 exon of wnk1 were associated with hsanii, the etiology of a substantial number of cases remains unexplained. in addition, the functions of wnk1/hsn2 and fam134b and their role in the peripheral nervous system remain poorly understood. using a yeast two-hybrid screen, ...201121820098
cryptococcal titan cell formation is regulated by g-protein signaling in response to multiple stimuli.the titan cell is a recently described morphological form of the pathogenic fungus cryptococcus neoformans. occurring during the earliest stages of lung infection, titan cells are 5-10 times larger than the normal yeast-like cells, thereby resisting engulfment by lung phagocytes and favoring persistence of infection. these enlarged cells exhibit altered capsule structure, thickened cell wall, increased ploidy, and resistance to nitrosative and oxidative stress. we demonstrate that two g protein- ...201121821718
kif18b interacts with eb1 and controls astral microtubule length during mitosis.regulation of microtubule (mt) dynamics is essential for proper spindle assembly and organization. kinesin-8 family members are plus-end-directed motors that modulate plus-end mt dynamics by acting as mt depolymerases or as mt plus-end capping proteins. in this paper, we show that the human kinesin-8 kif18b functions during mitosis to control astral mt organization. kif18b is a mt plus-tip-tracking protein that localizes to the nucleus in interphase and is enriched at astral mt plus ends during ...201121737685
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