Molecular analysis of the Saccharomyces cerevisiae YHR076w gene

Our results demonstrate that open reading frame (ORF) YHR076w on chromosome VIII of Saccharomyces cerevisiae that was previously described as a hypothetical gene is expressed. This ORF is transcribed as an mRNA of approximately 1,100 nucleotides. A 41.2-kDa polypeptide and three others predicted to be modified forms of Yhr076wp are detected by Western blot with a Yhr076wp-specific antibody. Promoter activity assays indicate that YHR076w transcription is regulated by carbon source and primarily by ethanol. Consistent with this observation, we have identified two potential ADR1 regulatory elements in the YHR076w upstream DNA region. Potential YAP1 and HSP elements are also identified in this region, suggesting other forms of regulation. YHR076w knockout strains do not exhibit any measurable growth or morphological phenotype under any conditions tested. However, increased YHR076w gene dosage confers a growth advantage to both wild-type and YHR076w knockout strains on 2% ethanol or 2% galactose medium in a low O2 growth environment. The fluorescence emitted by a Yhr076wp protein fusion to A. aquorin GFP colocalizes with the mitochondria in vivo.