In vivo evidence for non-universal usage of the codon CUG in Candida maltosa.
An alkane-assimilating yeast Candida maltosa had been studied in order to establish systems suitable for biotransformation of hydrophobic compounds. However, functional expression of heterologous genes tested for this purpose had not been successful in several cases. On the other hand, it had been reported that the codon CUG, a universal leucine codon, is read as serine in C. cylindracea. The same altered codon usage had also been suggested by in vitro experiments in some Candida yeasts which are phylogenetically closely related to C. maltosa. In this study we have shown that the failure in functional expression of a heterologous gene is due to the fact that the codon CUG is read as serine in C. maltosa. This conclusion was drawn from the following experimental results: (1) when a cytochrome P450 gene of C. maltosa containing a CTG codon was expressed in C. maltosa, the corresponding amino acid was found to be serine, and not leucine; (2) a tRNA gene with an almost identical structure to that of the tRNASerCAG gene of C. albicans could be isolated from the genome of C. maltosa; (3) the Saccharomyces cerevisiae URA3 gene, which has one CTG codon, could not complement the ura3 mutation of C. maltosa as itself, but when the CTG codon was changed to another leucine codon, CTC, the mutated gene could complement the ura3 mutation. The last result is the first example of succeeding in functional expression of a heterologous gene in Candida species having an altered codon usage by changing the CTG codon in the gene to another codon.
Sugiyama
H
H
Department of Agricultural Chemistry, University of Tokyo, Japan.
Ohkuma
M
M
Masuda
Y
Y
Park
S M
SM
Ohta
A
A
Takagi
M
M
D000595:Amino Acid Sequence; D000926:Anticodon; D001483:Base Sequence; D005075:Biological Evolution; D002175:Candida / Q000235:genetics*; D003062:Codon / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005800:Genes, Fungal; D005815:Genetic Code / Q000235:genetics; D007930:Leucine / Q000235:genetics; D008969:Molecular Sequence Data; D017354:Point Mutation / Q000235:genetics; D012362:RNA, Transfer, Ser / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017421:Sequence Analysis; D017423:Sequence Analysis, RNA; D012694:Serine / Q000235:genetics
D000926:Anticodon; D003062:Codon; D012362:RNA, Transfer, Ser; D012694:Serine; D003577:Cytochrome P-450 Enzyme System; D007930:Leucine