A deviation from the universal genetic code in Candida maltosa and consequences for heterologous expression of cytochromes P450 52A4 and 52A5 in Saccharomyces cerevisiae
- 29 January 1995
- Vol. 11 (1) , 33-41
- https://doi.org/10.1002/yea.320110105
Abstract
We demonstrate that serine instead of leucine is specified by the CUG codon in the yeast Candida maltosa. Evidence for this deviation from the universal genetic code was obtained by means of in vitro translation experiments. Depending on the cell‐free system used, either serine, in the C. maltosa system, or leucine, in the control with the conventional wheat germ system, was found to be incorporated into the translation products of artificial CUG‐containing mRNAs. Moreover, we were able to transfer the non‐universal decoding of CUG to the wheat germ system by adding a tRNA fraction isolated from C. maltosa. This finding indicates the presence in C. maltosa of an unusual serine tRNA that recognizes CUG. As a consequence of the altered genetic code, expression in Saccharomyces cerevisiae of C. maltosa cytochrome P450 genes required an exchange of their CTG triplets by TCT encoding serine in order to produce the authentic proteins. In contrast, heterologous expression of the original C. maltosa genes resulted in the formation of still active but unstable enzymes probably subject to selective proteolysis in the host cells.Keywords
This publication has 27 references indexed in Scilit:
- Evolutionary Position ofn-Alkane-assimilating YeastCandida maltosaShown by Nucleotide Sequence of Small-subunit Ribosomal RNA GeneBioscience, Biotechnology, and Biochemistry, 1993
- The P450 Superfamily: Update on New Sequences, Gene Mapping, Accession Numbers, Early Trivial Names of Enzymes, and NomenclatureDNA and Cell Biology, 1993
- Identification and Characterization of Additional Members of the Cytochrome P450 Multigene FamilyCYP52ofCandida tropicalisDNA and Cell Biology, 1992
- CYP52 (Cytochrome P450alk) Multigene Family inCandida maltosa: Molecular Cloning and Nucleotide Sequence of the Two Tandemly Arranged GenesDNA and Cell Biology, 1991
- Evidence That More Than One Gene Encodes n-Alkane-inducible Cytochrome P-450s in Candida maltosa, Found by Two-Step Gene Disruption.Agricultural and Biological Chemistry, 1991
- The codon CUG is read as serine in an asporogenic yeast Candida cylindraceaNature, 1989
- Molecular cloning and characterization of the primary structure of the alkane hydroxylating cytochrome P-450 from the yeast Candida maltosaBiochemical and Biophysical Research Communications, 1989
- Characterization of the alkane-inducible cytochrome P450 (P450alk) gene from the yeast Candida tropicalis: identification of a new P450 gene familyGene, 1989
- Purification of cytochrome P-450alk from n-alkane-grown cells of Candida maltosa, and cloning and nucleotide sequencing of the encoding gene.Agricultural and Biological Chemistry, 1989
- N-Alkanes induce the synthesis of cytochrome P-450 mRNA in Candida maltosaBiochemical and Biophysical Research Communications, 1986