The Arabidopsis thaliana proton transporters, AtNhx1 and Avp1, can function in cation detoxification in yeast.
AuthorsGaxiola RA, Rao R, Sherman A, Grisafi P, Alper SL, Fink GR
JournalProc Natl Acad Sci U S A
PubMed ID9990049
Overexpression of the Arabidopsis thaliana vacuolar H+-pyrophosphatase (AVP1) confers salt tolerance to the salt-sensitive ena1 mutant of Saccharomyces cerevisiae. Suppression of salt sensitivity requires two ion transporters, the Gef1 Cl- channel and the Nhx1 Na+/H+ exchanger. These two proteins colocalize to the prevacuolar compartment of yeast and are thought to ... More
The DExH box protein Suv3p is a component of a yeast mitochondrial 3'- to-5' exoribonuclease that suppresses group I intron toxicity.
AuthorsMargossian SP, Li H, Zassenhaus HP, Butow RA
JournalCell
PubMed ID8565066
The yeast mitochondrial protein Suv3p is a putative NTP-dependent RNA helicase. Here we report that in cells lacking Suv3p, there is an approximately 50-fold increase in the excised form of the group I intron omega of the mitochondrial 31S rRNA gene. Surprisingly, little mature 21S rRNA accumulates in those cells; ... More
A mouse homolog of the Escherichia coli recA and Saccharomyces cerevisiae RAD51 genes.
AuthorsMorita T, Yoshimura Y, Yamamoto A, Murata K, Mori M, Yamamoto H, Matsushiro A
JournalProc Natl Acad Sci U S A
PubMed ID8341671
'Analysis of mitotic and meiotic recombination in mammalian cells has been hampered by the complexity of the reactions involved as well as lack of mutants. Furthermore, none of the genes involved in the process has yet been identified. In budding yeast, Saccharomyces cerevisiae, the RAD51 gene is essential along with ... More
SALSA, a variant of yeast SAGA, contains truncated Spt7, which correlates with activated transcription.
Authors Sterner David E; Belotserkovskaya Rimma; Berger Shelley L;
JournalProc Natl Acad Sci U S A
PubMed ID12186975
'Spt-Ada-Gcn5 acetyltransferase (SAGA) is a previously described histone acetyltransferase/transcriptional coactivator complex in yeast. At promoters of certain genes (HIS3 and TRP3), SAGA has an inhibitory function involving a nonproductive TATA-binding protein interaction mediated by the Spt3 and Spt8 subunits. Related to this, Spt8-less SAGA is a major form of the ... More
Proteins connecting the nuclear pore complex with the nuclear interior.
AuthorsStrambio-de-Castillia C, Blobel G, Rout MP
JournalJ Cell Biol
PubMed ID10085285
'While much has been learned in recent years about the movement of soluble transport factors across the nuclear pore complex (NPC), comparatively little is known about intranuclear trafficking. We isolated the previously identified Saccharomyces protein Mlp1p (myosin- like protein) by an assay designed to find nuclear envelope (NE) associated proteins ... More
Functional expression of the murine Golgi CMP-sialic acid transporter in saccharomyces cerevisiae.
AuthorsBerninsone P, Eckhardt M, Gerardy-Schahn R, Hirschberg CB
JournalJ Biol Chem
PubMed ID9139716
'We have functionally expressed the murine Golgi putative CMP-sialic acid transporter in Saccharomyces cerevisiae. Using a galactose- inducible expression system, S. cerevisiae vesicles were able to transport CMP-sialic acid. Transport was dependent on galactose induction and was temperature-dependent and saturable with an apparent Km of 2.9 microM. Transport was inhibited ... More
Studies on identifying the catalytic role of Glu-204 in the active site of yeast invertase.
AuthorsReddy A, Maley F
JournalJ Biol Chem
PubMed ID8662946
'In a previous study on yeast invertase (Reddy, A., and Maley, F. (1990) J. Biol. Chem. 265, 10817-10820), we identified Asp-23 through the procedures of affinity labeling and site-directed mutagenesis as a catalytic nucleophile. In the present study we undertook to determine other residues involved in the catalytic process. Earlier ... More
Isolation and characterization of a thiamin transport gene, THI10, from Saccharomyces cerevisiae.
AuthorsEnjo F, Nosaka K, Ogata M, Iwashima A, Nishimura H
JournalJ Biol Chem
PubMed ID9235906
'We isolated a thiamin transporter gene, THI10, from a genomic library of Saccharomyces cerevisiae by the complementation of a yeast mutant defective in thiamin transport activity. The THI10 gene contained an open reading frame of 1,794 base pairs encoding a 598-amino acid polypeptide with a calculated molecular weight of 66, ... More
Significance of GTP hydrolysis in Ypt1p-regulated endoplasmic reticulum to Golgi transport revealed by the analysis of two novel Ypt1-GAPs.
Authors De Antoni Anna; Schmitzová Jana; Trepte Hans-Heinrich; Gallwitz Dieter; Albert Stefan;
JournalJ Biol Chem
PubMed ID12189143
'We here report on the identification and detailed biochemical characterization of two novel GTPase-activating proteins, Gyp5p and Gyp8p, whose efficient substrate is Ypt1p, a Ypt/Rab-GTPase essential for endoplasmic reticulum-to-Golgi trafficking in yeast. Gyp5p accelerated the intrinsic GTPase activity of Ypt1p 4.2 x 10(4)-fold and, surprisingly, the 40-fold reduced GTP hydrolysis ... More
Recombination-mediated PCR-directed plasmid construction in vivo in yeast.
AuthorsKevin R. Oldenburg + , Kham T. Vo , Susan Michaelis 1 and Chris Paddon*
JournalNucleic Acids Res
PubMed ID9016579
'We have extended the technique of PCR-directed recombination in Saccharomyces cerevisiae to develop a simple method for plasmid or gene construction in the absence of suitable restriction sites. The DNA to be cloned is PCR-amplified with 30-40 bp of homology to a linearized yeast plasmid. Co-transformation into yeast results in ... More
Cryopreserved yeast cells suitable for routine, small-scale transformations.
AuthorsTan SL, Dossett M, Katze MG
JournalBiotechniques
PubMed ID9821579
No abstract available.
In Saccharomyces cerevisiae, the inositol polyphosphate kinase activity of Kcs1p is required for resistance to salt stress, cell wall integrity, and vacuolar morphogenesis.
Authors Dubois Evelyne; Scherens Bart; Vierendeels Fabienne; Ho Melisa M W; Messenguy Francine; Shears Stephen B;
JournalJ Biol Chem
PubMed ID11956213
A problem for inositol signaling is to understand the significance of the kinases that convert inositol hexakisphosphate to diphosphoinositol polyphosphates. This kinase activity is catalyzed by Kcs1p in the yeast Saccharomyces cerevisiae. A kcs1Delta yeast strain that was transformed with a specifically ... More
Highly stereoselective reagents for beta-keto ester reductions by genetic engineering of baker's yeast.
Authors RodrÃguez S; Kayser M M; Stewart J D;
JournalJ Am Chem Soc
PubMed ID11456752
While whole cells of baker's yeast (Saccharomyces cerevisiae) are a convenient biocatalytic reducing agent for a wide variety of carbonyl compounds, mixtures of stereoisomeric alcohols are often observed since the organism contains a large number of reductase enzymes with overlapping substrate specificities but differing stereoselectivities. We sought to improve the ... More
Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and recA [published erratum appears in Nat Genet 1993 Nov;5(3):312]
AuthorsShinohara A, Ogawa H, Matsuda Y, Ushio N, Ikeo K, Ogawa T
JournalNat Genet
PubMed ID8358431
Rad51, of Saccharomyces cerevisiae, is a homologue of recA of Escherichia coli and plays crucial roles in both mitotic and meiotic recombination and in repair of double-strand breaks of DNA. We have cloned genes from human, mouse and fission yeast that are homologous to rad51. The 339 amino acid proteins ... More
Hsp90 binds and regulates Gcn2, the ligand-inducible kinase of the alpha subunit of eukaryotic translation initiation factor 2.
AuthorsDonze O, Picard D
JournalMol Cell Biol
PubMed ID10567567
Corrected] [published erratum appears in Mol Cell Biol 2000 Mar;20(5):1897The protein kinase Gcn2 stimulates translation of the yeast transcription factor Gcn4 upon amino acid starvation. Using genetic and biochemical approaches, we show that Gcn2 is regulated by the molecular chaperone Hsp90 in budding yeast Saccharomyces cerevisiae. Specifically, we found that ... More
An acyltransferase catalyzing the formation of diacylglucose is a serine carboxypeptidase-like protein
AuthorsLi AX, Steffens JC
JournalProc Natl Acad Sci U S A
PubMed ID10829071
1-O-beta-acyl acetals serve as activated donors in group transfer reactions involved in plant natural product biosynthesis and hormone metabolism. However, the acyltransferases that mediate transacylation from 1-O-beta-acyl acetals have not been identified. We report the identification of a cDNA encoding a 1-O-beta-acylglucose-dependent acyltransferase functioning in glucose polyester biosynthesis by Lycopersicon ... More
Conservation in budding yeast of a kinase specific for SR splicing factors.
AuthorsSiebel CW, Feng L, Guthrie C, Fu XD
JournalProc Natl Acad Sci U S A
PubMed ID10318902
SR protein kinases (SRPKs) and their substrates, the SR family of serine/arginine-rich pre-mRNA splicing factors, appear to be key regulators of alternative splicing. Although SR proteins have been well characterized through biochemical experiments in metazoans, their functions in vivo are unclear. Because of the strict splice site consensus and near ... More
B-Cell coactivator OBF-1 exhibits unusual transcriptional properties and functions in a DNA-bound Oct-1-dependent fashion.
AuthorsKrapp A, Strubin M
JournalMol Cell Biol
PubMed ID10330165
Eukaryotic transcriptional activators generally comprise both a DNA- binding domain that recognizes specific cis-regulatory elements in the target genes and an activation domain which is essential for transcriptional stimulation. Activation domains typically behave as structurally and functionally autonomous modules that retain their intrinsic activities when directed to a promoter by ... More
An oxidase-permease-based iron transport system in Schizosaccharomyces pombe and its expression in Saccharomyces cerevisiae.
AuthorsAskwith C, Kaplan J
JournalJ Biol Chem
PubMed ID8995275
Genetic studies have demonstrated that high affinity ferrous transport in Saccharomyces cerevisiae requires an oxidase (Fet3p) and a permease (Ftr1p). Using an iron-independent galactose-based expression system, we show that expression of these two genes can mediate high affinity ferrous iron transport, indicating that these two genes are not only necessary, ... More
DNA-protein interactions at the S.cerevisiae alpha 2 operator in vivo.
AuthorsMurphy MR, Shimizu M, Roth SY, Dranginis AM, Simpson RT
JournalNucleic Acids Res
PubMed ID8341604
Two homodimeric proteins, alpha 2 and MCM1, are required to repress transcription of a-cell type specific genes in haploid yeast alpha- cells. In vitro studies by others of the interactions of these proteins with operator DNA have suggested that MCM1 binds to the middle and alpha 2 to the ends ... More
An essential yeast protein, CBF5p, binds in vitro to centromeres and microtubules.
Yeast centromere DNA (CEN) affinity column chromatography has been used to purify several putative centromere and kinetochore proteins from yeast chromatin extracts. The single yeast gene (CBF5) specifying one of the major low-affinity centromere-binding proteins (p64'/CBF5p) has been cloned and shown to be essential for viability of Saccharomyces cerevisiae. CBF5 ... More
Identification of a novel membrane transporter associated with intracellular membranes by phenotypic complementation in the yeast Saccharomyces cerevisiae.
AuthorsHogue DL, Ellison MJ, Young JD, Cass CE
JournalJ Biol Chem
PubMed ID8621662
A partial mouse cDNA was isolated by its ability to functionally complement a thymidine transport deficiency in plasma membranes of the yeast, Saccharomyces cerevisiae. The full-length cDNA encoded a previously unidentified 27-kDa protein (mouse transporter protein (MTP)) with four predicted transmembrane-spanning domains. MTP mRNA was detected in cells of several ... More
Resolution of subunit interactions and cytoplasmic subcomplexes of the yeast vacuolar proton-translocating ATPase.
AuthorsTomashek JJ, Sonnenburg JL, Artimovich JM, Klionsky DJ
JournalJ Biol Chem
PubMed ID8626613
The vacuolar proton-translocating ATPase is the principal energization mechanism that enables the yeast vacuole to perform most of its physiological functions. We have undertaken an examination of subunit- subunit interactions and assembly states of this enzyme. Yeast two- hybrid data indicate that Vma1p and Vma2p interact with each other and ... More
The deduced sequence of the novel protransglutaminase E (TGase3) of human and mouse.
AuthorsKim IG, Gorman JJ, Park SC, Chung SI, Steinert PM
JournalJ Biol Chem
PubMed ID8099584
At least three transglutaminases are involved in terminal differentiation events in the epidermis and its derivatives, such as the hair follicle, presumably in cross-linking structural proteins and in the formation of the cornified cell envelope. Of these, only the transglutaminase 3 is a proenzyme, requiring activation by proteolytic cleavage, and ... More
Control of PKR protein kinase by hepatitis C virus nonstructural 5A protein: molecular mechanisms of kinase regulation.
AuthorsGale M Jr, Blakely CM, Kwieciszewski B, Tan SL, Dossett M, Tang NM, Korth MJ, Polyak SJ, Gretch DR, Katze MG
JournalMol Cell Biol
PubMed ID9710605
The PKR protein kinase is a critical component of the cellular antiviral and antiproliferative responses induced by interferons. Recent evidence indicates that the nonstructural 5A (NS5A) protein of hepatitis C virus (HCV) can repress PKR function in vivo, possibly allowing HCV to escape the antiviral effects of interferon. NS5A presents ... More
Yeast flavin-containing monooxygenase is induced by the unfolded protein response.
AuthorsSuh JK, Robertus JD
JournalProc Natl Acad Sci
PubMed ID10618381
Flavin-containing monooxygenase from yeast (yFMO) carries out the O(2)- and NADPH-dependent oxidation of biological thiols, including oxidizing glutathione to glutathione disulfide. FMO provides a large fraction of the oxidizing necessary for proper folding of disulfide bond-containing proteins; deletion of the enzyme reduces proper folding of endogenous carboxypeptidase Y by about ... More
Regulation of plant growth by cytokinin.
Authors Werner T; Motyka V; Strnad M; Schmülling T;
JournalProc Natl Acad Sci U S A
PubMed ID11504909
Cytokinins are a class of plant-specific hormones that play a central role during the cell cycle and influence numerous developmental programs. Because of the lack of biosynthetic and signaling mutants, the regulatory roles of cytokinins are not well understood. We genetically engineered cytokinin oxidase expression in transgenic tobacco plants to ... More
A Saccharomyces cerevisiae gene required for heterologous fatty acid elongase activity encodes a microsomal beta-keto-reductase.
A number of Saccharomyces cerevisiae membrane-bound oxidoreductases were examined for potential roles in microsomal fatty acid elongation, by assaying heterologous elongating activities in individual deletion mutants. One yeast gene, YBR159w, was identified as being required for activity of both the Caenorhabditis elegans elongase PEA1 (F56H11.4) and the Arabidopsis thaliana elongase ... More
A Bichaperone (Hsp70-Hsp78) System Restores Mitochondrial DNA Synthesis following Thermal Inactivation of Mip1p Polymerase.
Mitochondrial DNA synthesis is a thermosensitive process in the yeast Saccharomyces cerevisiae. We found that restoration of mtDNA synthesis following heat treatment of cells is dependent on reactivation of the mtDNA polymerase Mip1p through the action of a mitochondrial bichaperone system consisting of the Hsp70 system and the Hsp78 oligomeric ... More
Mitochondrial protein import: molecular basis of the ATP-dependent interaction of MtHsp70 with Tim44.
Protein translocation across the mitochondrial inner membrane is driven by cycles of binding and release of mitochondrial heat shock protein 70 (mtHsp70) in the matrix. The peripheral inner membrane protein, Tim44, recruits mtHsp70 in an ATP-dependent manner to the import sites. We show that DnaK, the closely related Hsp70 of ... More
A vertebrate fatty acid desaturase with Delta 5 and Delta 6 activities.
Authors Hastings N; Agaba M; Tocher D R; Leaver M J; Dick J R; Sargent J R; Teale A J;
JournalProc Natl Acad Sci U S A
PubMed ID11724940
Delta5 and Delta6 fatty acid desaturases are critical enzymes in the pathways for the biosynthesis of the polyunsaturated fatty acids arachidonic, eicosapentaenoic, and docosahexaenoic acids. They are encoded by distinct genes in mammals and Caenorhabditis elegans. This paper describes a cDNA isolated from zebrafish (Danio rerio) with high similarity to ... More
Mutagenesis of a functional chimeric gene in yeast identifies mutations in the simian virus 40 large T antigen J domain.
Authors Fewell Sheara W; Pipas James M; Brodsky Jeffrey L;
JournalProc Natl Acad Sci U S A
PubMed ID11854498
Simian virus 40 large T antigen contains an amino terminal J domain that catalyzes T antigen-mediated viral DNA replication and cellular transformation. To dissect the role of the J domain in these processes, we exploited the genetic tools available only in the yeast Saccharomyces cerevisiae to isolate 14 loss-of-function point ... More
Identification and characterization of a sphingolipid delta 4-desaturase family.
Sphingolipids desaturated at the Delta4-position are important signaling molecules in many eukaryotic organisms, including mammals. In a bioinformatics approach, we now identified a new family of protein sequences from animals, plants, and fungi and characterized these sequences biochemically by expression in Saccharomyces cerevisiae. This resulted in the identification of the ... More
Glucosylceramide synthases, a gene family responsible for the biosynthesis of glucosphingolipids in animals, plants, and fungi.
Authors Leipelt M; Warnecke D; Zähringer U; Ott C; Müller F; Hube B; Heinz E;
JournalJ Biol Chem
PubMed ID11443131
Glucosylceramides are membrane lipids in most eukaryotic organisms and in a few bacteria. The physiological functions of these glycolipids have only been documented in mammalian cells, whereas very little information is available of their roles in plants, fungi, and bacteria. In an attempt to establish appropriate experimental systems to study ... More
Functional activity and role of cation-efflux family members in Ni hyperaccumulation in Thlaspi goesingense.
Authors Persans M W; Nieman K; Salt D E;
JournalProc Natl Acad Sci U S A
PubMed ID11481436
The ability of Thlaspi goesingense to hyperaccumulate Ni seems to be governed in part by enhanced accumulation of Ni within leaf vacuoles. We have characterized genes from T. goesingense encoding putative vacuolar metal ion transport proteins, termed metal tolerance proteins (TgMTPs). These proteins contain all of the features of cation-efflux ... More