RESUMO
We have characterized the KlROX1 gene from Kluyveromyces lactis and verified that it does not regulate the hypoxic response in this yeast, oppositely to the Saccharomyces cerevisiae homologue ScROX1. The KlROX1 promoter is not regulated by KlHap1p or KlRox1p in response to changes aerobiosis/hypoxia. Besides, KlRox1p expression only partially represses ScANB1 in S. cerevisiae and does not regulate the ScANB1 and KlHEM13 promoters in K. lactis. KlRox1p does not interact either with KlTup1p or KlSsn6p or with their homologues ScTup1p and ScSsn6p, which are components of the general co-repressor factor that mediates the transcriptional repression exerted by ScRox1p in S. cerevisiae. We have found that KlROX1 mediates the response to arsenate and cadmium and, in the presence of cadmium, it is necessary for KlYCF1 expression, a gene encoding a protein with homology to the yeast cadmium and arsenite vacuolar transporter. EMSA assays show that KlRox1p binds, through its HMG domain, to a DNA sequence present in the KlYCF1 promoter. Although in S. cerevisiae the function of ScRox1p in cadmium resistance was already known and linked to regulation of ScFET4 expression, we have found that ScRox1p also regulates ScYCF1transcription and binds to its promoter.
