Isolation and sequence analysis of the gene encoding translation elongation factor 3 from Candida albicans

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Abstract
The structural gene encoding translation elongation factor 3 (EF‐3) has been cloned from a Candida albicans genomic library by hybrization to a Saccharomyces cerevisiae probe containing the Saccharomyces gene, YEF3 (Sandbaken et al., 1990b). The sequences were shown to be functionally homologous to the Saccharamyces gene by three criteria: (1) a Saccharomyces strain transformed with a high copy plasmid containing CaEF3 sequences 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) 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 (Sandaken et 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 among the two organisms.