Molecular characterization of MRG19 of Saccharomyces cerevisiae. Implication in the regulation of galactose and nonfermentable carbon source utilization.
We have reported previously that multiple copies of MRG19 suppress GAL genes in a wild-type but not in a gal80 strain of Saccharomyces cerevisiae. In this report we show that disruption of MRG19 leads to a decrease in GAL induction when S. cerevisiae is induced with 0.02% but not with 2.0% galactose. Disruption of MRG19 in a gal3 background (this strain shows long-term adaptation phenotype) further delays the GAL induction, supporting the notion that its function is important only under low inducing signals. As a corollary, disruption of MRG19 in a gal80 strain did not decrease the constitutive expression of GAL genes. These results suggest that MRG19 has a role in GAL regulation only when the induction signal is weak. Unlike the effect on GAL gene expression, disruption of MRG19 leads to de-repression of CYC1-driven beta-galactosidase activity. MRG19 disruptant also showed a twofold increase in the rate of oxygen uptake as compared with the wild-type strain. ADH2, CTA1, DLD1, and CYC7 promoters that are active during nonfermentative growth did not show any de-repression of beta-galactosidase activity in the MRG19 disruptant. Western blot analysis indicated that MRG19 is a glucose repressible gene and is expressed in galactose and glycerol plus lactate. Experiments using green fluorescent protein fusion constructs indicate that Mrg19p is localized in the nucleus consistent with the presence of a consensus nuclear localization signal sequence. Based on the above results, we propose that Mrg19p is a regulator of galactose and nonfermentable carbon utilization.
Khanday
Firdous A
FA
Laboratory of Molecular Genetics, Biotechnology Center, Indian Institute of Technology, Powai, Mumbai, India.
Saha
Maitreyi
M
Bhat
Paike Jayadeva
PJ
D001483:Base Sequence; D015153:Blotting, Western; D002244:Carbon / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D045304:Cytochromes c; D017931:DNA Primers; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics* / Q000502:physiology; D005690:Galactose / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
C014059:CYC1 protein, S cerevisiae; D003574:Cytochrome c Group; D017931:DNA Primers; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D002244:Carbon; D045304:Cytochromes c; D005690:Galactose