摘要:

AbstractThe structural gene encoding translation elongation factor 3 (EF‐3) has been cloned from aCandida albicansgenomic library by hybrization to aSaccharomyces cerevisiaeprobe containing theSaccharomycesgene,YEF3(Sandbakenet al., 1990b). The sequences were shown to be functionally homologous to theSaccharamyces geneby three criteria: (1) aSaccharomycesstrain transformed with a high copy plasmid containingCaEF3sequences overprodues the EF‐3 peptide two‐fold; (2) extracts from this strain exhibit a two‐fold increase in the Ef‐3 catalysed, ribosome‐dependent ATPase activity (Kamath and Chakraburtty, 1988); and (3) theCandidagene complements aSaccharomycesnull mutant. The coding region, identified by DNA sequencing, indicates thatCaEF3encodes a 1050 amino acid polypeptide having a potential molecular weight of 116 865 Da. This protein shows 77% overall identity to theSaccharomyces YEF3gene, with a significantly greater identity (94%) concentrated in the region of the protein thought to contain the catalytic domain of EF‐3 (Sandakenet al., 1990a). The upstream non‐coding region contains T‐rich regions typical of many yeast genes and several potential RAPI/GRFI elements shown to regulate expression of a number of translational genes (Mager, 1988). The data confirm a high degree of conservation for EF‐3 amon

ISSN:0749-503X    

DOI:10.1002/yea.320080502

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractProteinase B precursors are modified by an N‐linked carbohydrate side chain at Asn 314. Glycosylation at this position is not required for proper localization, processing, or activation of the enzym

ISSN:0749-503X    

DOI:10.1002/yea.320080503

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractA strain of the methylotrophic yeastHansenula polymorpha, A16 has been developed that expresses the guar α‐galactosidase gene to 22.4 mg/g dry cell weight in chemostat cultures at a dilution rate of 0.1 h−1. This corresponds to more than 13.1% of solube cell protein, of which 56–62% is secreted into the medium. The α‐galactosidase gene was flanked by the promoter and terminator sequences of theH. polymorpha moxgene, which can direct expression of themoxgene itself more than 30% of total cell protein under methanol growth. The expression cassette (pUR3510) based on theSaccharomyces cerevisiaeplasmid, YEp13, was integrated into the genome. Such transformants were stable in chemostat cultures and exhibited 100% stability for both α‐galactosidase+and leu+phenotypes. Chemostat cultures produced higher levels of α‐galactosidase with higher specific productive expressed as mg α‐galactosidase g−1h−1comp

ISSN:0749-503X    

DOI:10.1002/yea.320080504

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractThe killer phenotype expressed bySaccharomyces cerevisiaestrain 28 differs fron that of the more extensively studied K1and K2killers with respect to immunity, mode of toxin action and cell wall primary toxin receptor. We previosly demonstrated that the M28and L28dsRNAs found in strain 28 are present in virus‐like particles (VLPs) and that transfection with these VLPs is sufficient to confer the complete K28phenotype on a dsRNA‐free recipient cell. We also demonstrated that L28, like the L‐A‐H species in K1killers, has [HOK] activity required for maintenance of M1‐dsRNA, and predicted that M28would share with M1dependence on L‐A for replication. We now confirm this prediction by genetic and biochemical analysis of the effects of representativemak,skiandmktmutations on M28maintenance, demonstrating that M28replication resebles M1in all respects. We also show that L28is an L‐A‐H species lacking [B] activity, and that M28excludes both M1and M2from the same cytoplasm. Stable coexpression of K28phenotype from M28and of K1phenotype from an M1‐cDNA clone was demonstrated. Exclusion, therefore, acts at the level of dsRNA replication, presumably reflecting competition for the L‐A‐H encoded capsid and cap‐pol fusion protein, rather than reflecting incompatibility of toxin or immunity expression. Finally, we show that expression of active K28toxin, bu t not of K28immunity, require

ISSN:0749-503X    

DOI:10.1002/yea.320080505

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractTheRAD54gene ofSaccharomyces cerevisiaeis involved in the recombinational repair of DNA damage. The predicted amino acid sequence of the RAD54 protein shows significant homologies with the yeast SNF2 protein, which is required for the transcriptioal activation of a number of diversely regulated genes. These proteins are 31% identical in a 492‐amino acid region that includes presumed nucleotide and Mg2+binding sites. We noted previously that the SNF2 protein also shares homology with a partial open reading frame (ORF) that was reported with the sequence of an adjacent gene. This ORF also shares homology with the RAD54 protein. To test whether this ORF is involved in transcriptional activation or DNA repair, yeast strains deleted for part of it have been isolated. These strains do not show a Snf‐like phenotyp, but they are UV sensitive. This gene has been identified asRAD 16, a gene involved in the excision repair of DNA damage. Analysis of therad16deletion mutations indicates thatRAD16encodes a nonessential function and is not absolutely required for excision repair. Outside the region of homology to RAD54 and SNF2, the predicted RAD16 protein contains a novel cysteine‐rich motif that may bind zinc and that has been found recently in eleven other proteins, including the yeast RAD18 protein. The homologies betweenRAD16, RAD54andSNF2are also shared by several additional, recently isolated yeast andDrosophila

ISSN:0749-503X    

DOI:10.1002/yea.320080506

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractWe have analysed a region some 30 kb centromere distal formPHO5on the right arm of yeast chromosome II and determined the nucleotide sequence of a 8.95 kb DNA segment from this region. By this analysis we were able to derive the precise location and the transcriptional orientation ofCMD1, ALG1, SSN6andLYS2.An open reading frame of 2370 bp was locatlized betweenSSN6andLYS2, which has recently been identified (Schildetal., 1991) to be theRAD16gene. The putative gene product, 790 amino acids in length, reveals several interesting freatures. It contains a nuclear target singnature and shares several blocks of similarity with the yeast recombinational repair protein RAD54 and the nuclear factor SNF2 (SW12), which is required for teh transcriptioal activation of a number of yeast genes. The similarity blocks in these three proteins are reminiscent of those found in the helicase superfamily. Furthrmore, RAD16 contains a novel ‘double‐finger’ motif, which has been encountered in a variety of proteins from different organisms that are suggested to interact with DNA and are involved in diverse functions including site‐specific recombination, DNA repair, and transcriptional regulation. The putative gene product ofRAD16then is the first example of a proteins in which the novel double‐finger motif is found to be combined with a poteintial DNA helicase

ISSN:0749-503X    

DOI:10.1002/yea.320080507

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractWe have entirely sequenced a 10,835 bp segment of the right arm from chromosome III contained in the J11D and J11D‐K3B GF clones. The segment contains seven open reading frames longer than 100 amino acids. Three of them,RVS161(Urdaciet al., 1990; Crouzetet al., 1991),ADP1(Purnelleet al., 1991) andPGK1, (Hitzemanet al., 1982) have been described previously.YCR10C, encodes a putative membrane protein.YCR8W, (encoding a putative protein kinase) andYCR14cextend inside the D10H (Skalaet al., 1991) and 62B5‐2D clones respectively. FourARSelements previously reported by Palzkillet al., (1986) are located betweenRVS161andYCR

ISSN:0749-503X    

DOI:10.1002/yea.320080508

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractWe have isolated and sequenced a novel gene fromSaccharomyces cerevisiae codingfor an essential protein of unknown function. The gene calledYJU2was borne on a chromosome X fragments shown by hybridization to intactS. cerevisiaechromosomal DNA fractionated by orthogonal pulsed field electrophoresis. Northern hybridization analysis indicated that the 0.8‐kb transcript ofYJU2is expressed in exponential‐phase cells grown in rich medium (data not shown). Figure 1 shows the nucleotide and deduced amino‐acid sequences of the 834‐bp coding region as well as the nucleotide sequences of the 5′ upstream region and of the 3′ downstream region, together with the flanking neighbouring open reading frames (ORFs),

ISSN:0749-503X    

DOI:10.1002/yea.320080509

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

ISSN:0749-503X    

DOI:10.1002/yea.320080501

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY