Isolation and sequence analysis of the gene encoding translation elongation factor 3 from Candida albicans.
The structural gene encoding translation elongation factor 3 (EF-3) has been cloned from a Candida albicans genomic library by hybridization to a Saccharomyces cerevisiae probe containing the Saccharomyces gene, YEF3 (Sandbaken et al., 1990b). The sequences were shown to be functionally homologous to the Saccharomyces gene by three criteria: (1) a Saccharomyces strain transformed with a high copy plasmid containing CaEF3 sequences overproduces 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) the Candida gene complements a Saccharomyces null mutant. The coding region, identified by DNA sequencing, indicates that CaEF3 encodes a 1050 amino acid polypeptide having a potential molecular weight of 116,865 Da. This protein shows 77% overall identity to the Saccharomyces YEF3 gene, with a significantly greater identity (94%) concentrated in the region of the protein thought to contain the catalytic domain of EF-3 (Sandbaken et al., 1990a). The upstream non-coding region contains T-rich regions typical of many yeast genes and several potential RAP1/GRF1 elements shown to regulate expression of a number of translational genes (Mager, 1988). The data confirm a high degree of conservation for EF-3 among the two organisms.
Di Domenico
B J
BJ
Schering-Plough Research Institute, Bloomfield, NJ 07003.
Lupisella
J
J
Sandbaken
M
M
Chakraburtty
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010445:Peptide Elongation Factors / Q000235:genetics*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D004271:DNA, Fungal; D005656:Fungal Proteins; D010445:Peptide Elongation Factors; D029701:Saccharomyces cerevisiae Proteins; C111584:YEF3 protein, S cerevisiae