S. Sakajo et al., Structure and regulatory expression of a single copy alternative oxidase gene from the yeast Pichia anomala, BIOS BIOT B, 63(11), 1999, pp. 1889-1894
To investigate the regulatory mechanism of alternative oxidase gene express
ion, genomic DNA was cloned from the yeast Pichia anomala. Genomic Southern
blot analysis suggested that a single copy nuclear gene encoded an alterna
tive oxidase in the yeast. The nucleotide sequence showed an uninterrupted
coding region for the alternative oxidase protein. In the upstream region f
rom the transcription initiation site found by primer extension analysis, C
CAAT, TATAA, and UAS2-like elements were detected. The UAS2 is the element
involved in transcriptional regulation by carbon source and the target site
for the factor, HAP2/3/4/5 protein complex, in Saccharomyces cerevisiae. B
y a gel mobility shift assay, a specific retardation band was detected when
a protein extract from cells grown on an inducing carbon source was incuba
ted with a UAS2-containing probe. These results suggest that carbon source
regulation of alternative oxidase gene expression is mediated by the UAS2-l
ike element and a MAP-like factor in P. anomala.