Expression of the Saccharomyces cerevisiae AAC2 gene encoding the major mitochondrial ADP/ATP carrier was examined. The intracellular level of the carrier protein, as well as the level of the AAC2-gene-specific mRNA, is influenced by the presence or absence of oxygen or of heme, and it is subject to carbon-source control. In addition, the expression of AAC2 gene requires the products of the HAP2 and HAP3 genes, but not that of the HAP1 gene. The 5'-flanking region of the gene was isolated, sequenced and fused to the lacZ reporter gene in order to study the effect of carbon sources and of specific deletion mutations on expression of the gene in yeast transformants. The expression of the reporter gene reveals that the AAC2 gene possesses a strong inducible promoter. The promoter analysis, combined with expression studies in the wild-type as well as in various mutant strains, identified an upstream activation site (UAS) contained within a sequence between -393 bp and -268 bp, and several major initiation sites of AAC2 mRNA between -105bp and -95 bp. Deletion analysis also shows that the TATA boxes located 45 bp and 104 bp upstream of the 5'-ends of AAC2 mRNA are not essential for the transcription. The UAS of the AAC2 gene is required for activation by HAP2 and heme and for release from glucose repressin. A restriction fragment containing the UAS conferred oxygen and carbon source regulation when placed upstream of another yeast gene encoding ADP/ATP carrier (AAC3), deleted of its regulatory sequences. The UAS of the AAC2 gene contains at least two distinct motifs for DNA-binding transcriptional activators, including one which is identical with the core HAP2/3/4 binding motif, and a second one with the ABF1 consensus binding sequence. Our results indicate that these sequences mediate the effects of the respective transactivator on the oxygen- and carbon-source-dependent transcription of the AAC2 gene.

• PMID: 7758459
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Journal Article | Research Support, Non-U.S. Gov't

Authors

Betina S, Gavurníková G, Haviernik P, Sabová L, Kolarov J

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