Life with 6000 Genes

Goffeau, André; Barrell, Bart; Bussey, Howard; Davis, Ronald W.; Dujon, Bernard; Feldmann, Horst; Galibert, Francis; Hoheisel, Jörg D.; Jacq, Claude; Johnston, Mark; Louis, Edward J.; Mewes, Hans‐Werner; Murakami, Yasufumi; Philippsen, Peter; Tettelin, Hervé; Oliver, Stephen G.

doi:10.1126/science.274.5287.546

Cited by 3,737 publications

(2,368 citation statements)

References 60 publications

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“…termini generated by hammerhead cleavage to serve as substrates for poly(A) polymerase (Tanner, 1999;Zhao et al+, 1999)+ In yeast, it has been possible to obtain polyadenylation of a ribozyme-containing transcript (Egli & Braus, 1994), an event that most likely depends on RNA sequence context, that is, triggering cryptic cleavage/polyadenylation signals+ Because we were unable to detect any polyadenylated transcripts in RACE experiments we conclude that the TRP4-ribozyme mRNA does not contain any cryptic signals+ Although mRNA 39-end processing and nuclear export have been shown to be linked (Eckner et al+, 1991;Hilleren et al+, 2001), the TRP4-ribozyme mRNA appears to be efficiently exported from the nucleus+ It is unclear whether this occurs by pathways previously shown to be effective for nonpolyadenylated RNAs like rRNA and tRNAs (Moy & Silver, 1999;Sarkar et al+, 1999;Simos & Hurt, 1999) or by some other mechanism+ Unlike metazoans and their unadenylated histone mRNAs, there are no known examples of yeast RNA polymerase II transcripts that lack poly(A) tails (Goffeau et al+, 1996)+ Future studies to assess the mechanism by which such nonpolyadenylated mRNAs are exported will be of interest+…”

Section: Discussionmentioning

confidence: 99%

Replacement of the yeast TRP4 3??? untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail

Düvel

Valerius

Mangus

et al. 2002

RNA

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The mRNA poly(A) tail serves different purposes, including the facilitation of nuclear export, mRNA stabilization, efficient translation, and, finally, specific degradation. The posttranscriptional addition of a poly(A) tail depends on sequence motifs in the 39 untranslated region (39 UTR) of the mRNA and a complex trans-acting protein machinery. In this study, we have replaced the 39 UTR of the yeast TRP4 gene with sequences encoding a hammerhead ribozyme that efficiently cleaves itself in vivo. Expression of the TRP4-ribozyme allele resulted in the accumulation of a nonpolyadenylated mRNA. Cells expressing the TRP4-ribozyme mRNA showed a reduced growth rate due to a reduction in Trp4p enzyme activity. The reduction in enzyme activity was not caused by inefficient mRNA export from the nucleus or mRNA destabilization. Rather, analyses of mRNA association with polyribosomes indicate that translation of the ribozyme-containing mRNA is impaired. This translational defect allows sufficient synthesis of Trp4p to support growth of trp4 cells, but is, nevertheless, of such magnitude as to activate the general control network of amino acid biosynthesis.

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Section: Discussionmentioning

confidence: 99%

Replacement of the yeast TRP4 3??? untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail

Düvel

Valerius

Mangus

et al. 2002

RNA

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“…P39743) that reduces viability upon starvation (Rvs167p; Bauer et al, 1993); ORF2 encodes a peptide homologous to S. cerevisiae Pob3p (POB3 : Accession No. NP 013642) that binds to the catalytic subunit of DNA polymerase-α (Pol1p) (Goffeau et al, 1996).…”

Section: Resultsmentioning

confidence: 99%

Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547

Wang

Kayingo

Blomberg

et al. 2002

Yeast

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The dihydroxyacetone pathway, an alternative pathway for the dissimilation of glycerol via reduction by glycerol dehydrogenase and subsequent phosphorylation by dihydroxyacetone (DHA) kinase, is activated in the yeasts Saccharomyces cerevisiae and Zygosaccharomyces rouxii during osmotic stress. In experiments aimed at investigating the physiological function of the DHA pathway in Z. rouxii, a typical osmotolerant yeast, we cloned and characterized a DAK gene encoding dihydroxyacetone kinase from Z. rouxii NRRL 2547. Sequence analysis revealed a 1761 bp open reading frame, encoding a peptide composed of 587 deduced amino acids with the predicted molecular weight of 61 664 Da. As the amino acid sequence was most closely homologous (68% identity) to the S. cerevisiae Dak1p, we named the gene and protein ZrDAK1 and ZrDak1p, respectively. A putative ATP binding site was also found but no consensus element associated with osmoregulation was found in the upstream region of the ZrDAK1 gene. The ZrDAK1 gene complemented a S. cerevisiae W303-1A dak1 dak2 strain by improving the growth of the mutant on 50 mmol/l dihydroxyacetone and by increasing the tolerance to dihydroxyacetone in a medium containing 5% sodium chloride, suggesting that it is a functional homologue of the S. cerevisiae DAK1. However, expression of the ZrDAK1 gene in the S. cerevisiae dak1 dak2 strain had no significant effect on glycerol levels during osmotic stress. The ZrDAK1 sequence has been deposited in the public data bases under Accession No. AJ294719; regions upstream and downstream of ZrDAK1 are deposited as Accession Nos AJ294739 and AJ294720, respectively.

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“…This is largely because S288C was the strain chosen for the yeast genome sequencing project (Goffeau et al, 1996). Winston et al (1995) carried out the first construction of a set of genetically marked strains by the direct manipulation of S288C, creating the FY series.…”

Section: Genetic Background Of Yeast Strainsmentioning

confidence: 99%

A new family of yeast vectors and S288C‐derived strains for the systematic analysis of gene function

Tomlin¹,

Wixon²,

Bolotin‐Fukuhara³

et al. 2001

Yeast

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The yeast genome has been shown to contain a significant number of gene families with more than three members. In order to study these families it is often necessary to generate strains carrying deletions of all members of the family, which can require a wide range of auxotrophic markers. To facilitate such studies, we have generated yeast strains containing deletions of a selection of nutritional marker genes (ade2, ade4, ade8, met3 and met14). We have also cloned the corresponding cognate genes, allowing their use in PCR-based gene disruptions. Two new pRS family Saccharomyces cerevisiae-Escherichia coli shuttle vectors containing ADE8 (one low-copy, pRS4110, and one high-copy, pRS4210) have been produced for use in conjunction with the new strains. A system for easier synthetic lethal screening using one of these new markers is also presented. The ADE8 and HIS3 genes have been cloned together on a high-copy vector (pRS4213), providing a plasmid for red-white colour screening in the ade2D0 ade8D0 strains we have generated. In contrast to some conventional systems, this plasmid allows for screening using gene libraries constructed in URA3 plasmids.

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Life with 6000 Genes

Cited by 3,737 publications

References 60 publications

Replacement of the yeast TRP4 3??? untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail

Replacement of the yeast TRP4 3??? untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail

Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547

A new family of yeast vectors and S288C‐derived strains for the systematic analysis of gene function

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