Isolation of the yeast gene encoding elongation factor 3 for protein synthesis.

Qin, Shiliang; Moldave, Kivie; McLaughlin, Calvin S.

doi:10.1016/s0021-9258(18)47639-9

Cited by 30 publications

(1 citation statement)

References 21 publications

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“…After prehybridization for 1 hr in hybridization buffer at the desired temperature, hybridization of the filters was carried out as described by Sundstrom and Sypherd (13). The 3.5 kb XbaI fragment from pYEF-3 (16) or the 981 bp EcoRI fragment from p3R4 (C. albicans genomic clone isolated in this work) were labelled (17,18) with [c&32P]dGTP (Dupont-NEN) using a random-primer kit (United States Biochemical) for use as hybridization probes. Hybridization was accomplished with 5 x 106cpm of labelled probe at temperatures of 37°C and 42°C for the S. cerevisiae and C. albicans probes, respectively.…”

Section: Methodsmentioning

confidence: 99%

Isolation and sequence analysis of the gene for translation elongation factor 3 fromCandida albicans

Myers

Fonzi

Sypherd³

1992

Nucl Acids Res

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Elongation factor 3 (EF-3) is a unique and essential component of the translational system in fungi. The gene, CEF-3, encoding elongation factor 3 has been isolated from the dimorphic fungus Candida albicans. A heterologous gene probe containing the coding region of the EF-3 gene from Saccharomyces cerevisiae (YEF-3) was used to screen three Candida albicans genomic DNA libraries. The nucleotide sequences of four partial clones were determined and combined for a full-length of 3,671 base pairs (bp). A continuous open reading frame (ORF) of 3,147 bp encoding a predicted protein of 1,049 amino acids and Mr of 116,739 daltons has been identified. A transcript of 3,400 nucleotides is seen in Northern blot hybridization of Candida albicans total RNA using a CEF-3 gene probe. The single locus CEF-3 gene maps to chromosome 5 in the genome. Comparison of the deduced amino acid sequences of CEF-3 and YEF-3 shows 77.6%. identity. A higher degree of identity, 86.5%, is found when comparing the carboxy-terminal portions of the two proteins. At the nucleotide level, comparison of the coding regions of the two genes exhibit 79% identity while the upstream and downstream regions show 46% and 40% identity, respectively.

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Section: Methodsmentioning

confidence: 99%

Isolation and sequence analysis of the gene for translation elongation factor 3 fromCandida albicans

Myers

Fonzi

Sypherd³

1992

Nucl Acids Res

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Identification of Genes with Nutrient-controlled Expression by PCR-mapping in the YeastSaccharomyces cerevisiae

Crauwels

Winderickx

Winde

et al. 1997

Yeast

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We have used RNA fingerprinting by the mRNA Differential Display technique to identify new genes in the yeast Saccharomyces cerevisiae, expression of which is controlled by specific nutrient conditions. mRNA was isolated from cells grown on glucose medium into exponential and stationary phase, and from cells starved for nitrogen on glucose‐containing medium. To avoid interference with the large number of glucose‐repressible genes, a glucose‐repression‐deficient strain was used. Twenty different sets of arbitrary primers chosen at random were used for PCR‐amplification of reverse transcriptase generated cDNAs, which resulted in six highly reproducible gene expression patterns. The validity of the approach was confirmed by sequencing PCR products of genes with known expression patterns, SUP44/RPS4, CTT1, SSA3, HSP30 and HSP104, and genes with related functions, TEF1 and TEF3, encoding translation elongation factors. In all cases the specificity of the responses was confirmed by Northern blot analysis. The results show that the PCR‐mapping method is highly useful for the identification of new genes expressed under specific conditions in the yeast S. cerevisiae © 1997 John Wiley & Sons, Ltd.

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Identification and kinetic analysis of a functional homolog of elongation factor 3,YEF3 inSaccharomyces cerevisiae

Sarthy

McGonigal

Capobianco

et al. 1998

Yeast

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Yeast and other fungi contain a soluble elongation factor 3 (EF‐3) which is required for growth and protein synthesis. EF‐3 contains two ABC cassettes, and binds and hydrolyses ATP. We identified a homolog of the YEF3 gene in the Saccharomyces cerevisiae genome database. This gene, designated YEF3B, is 84% identical in protein sequence to YEF3, which we will now refer to as YEF3A. YEF3B is not expressed during growth under laboratory conditions, and thus cannot rescue growth of YEF3A deletion strains. However, YEF3B can take the place of YEF3A in vivo when expressed from the YEF3A or ADH1 promoters. The products of the YEF3A and YEF3B genes, EF‐3A and EF‐3B, respectively, were expressed from the ADH1 promoter and purified. Both factors possessed basal and ribosomal‐stimulated ATPase activity, and had similar affinity for yeast ribosomes (103 to 113 nm). Km values for ATP were similar, but the Kcat values differed significantly. Ribosome‐dependent ATPase activity of EF‐3A was more efficient than EF‐3B, since the Kcat and Kcat/Km values for EF‐3A were about two‐fold higher; however, the difference in Kcat/Km values between the two factors was small for basal ATPase activity. © 1998 John Wiley & Sons, Ltd.

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Isolation of the yeast gene encoding elongation factor 3 for protein synthesis.

Cited by 30 publications

References 21 publications

Isolation and sequence analysis of the gene for translation elongation factor 3 fromCandida albicans

Isolation and sequence analysis of the gene for translation elongation factor 3 fromCandida albicans

Identification of Genes with Nutrient-controlled Expression by PCR-mapping in the YeastSaccharomyces cerevisiae

Identification and kinetic analysis of a functional homolog of elongation factor 3,YEF3 inSaccharomyces cerevisiae

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