Multiple phosphorylated forms of the Saccharomyces cerevisiae Mcm1 protein include an isoform induced in response to high salt concentrations.
The Saccharomyces cerevisiae Mcm1 protein is an essential multifunctional transcription factor which is highly homologous to human serum response factor. Mcm1 protein acts on a large number of distinctly regulated genes: haploid cell-type-specific genes, G2-cell-cycle-regulated genes, pheromone-induced genes, arginine metabolic genes, and genes important for cell wall and cell membrane function. We show here that Mcm1 protein is phosphorylated in vivo. Several (more than eight) isoforms of Mcm1 protein, resolved by isoelectric focusing, are present in vivo; two major phosphorylation sites lie in the N-terminal 17 amino acids immediately adjacent to the conserved MADS box DNA-binding domain. The implications of multiple species of Mcm1, particularly the notion that a unique Mcm1 isoform could be required for regulation of a specific set of Mcm1's target genes, are discussed. We also show here that Mcm1 plays an important role in the response to stress caused by NaCl. G. Yu, R. J. Deschenes, and J. S. Fassler (J. Biol. Chem. 270:8739-8743, 1995) showed that Mcm1 function is affected by mutations in the SLN1 gene, a signal transduction component implicated in the response to osmotic stress. We find that mcm1 mutations can confer either reduced or enhanced survival on high-salt medium; deletion of the N terminus or mutation in the primary phosphorylation site results in impaired growth on high-salt medium. Furthermore, Mcm1 protein is a target of a signal transduction system responsive to osmotic stress: a new isoform of Mcm1 is induced by NaCl or KCl; this result establishes that Mcm1 itself is regulated.
Kuo
M H
MH
Department of Biochemistry and Biophysics, School of Medicine, University of Rochester, New York 14642, USA.
Nadeau
E T
ET
Grayhack
E J
EJ
D000135:Acid Phosphatase; D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007525:Isoelectric Focusing; D018021:Lithium Chloride / Q000494:pharmacology; D026341:Minichromosome Maintenance 1 Protein; D008969:Molecular Sequence Data; D009154:Mutation; D009994:Osmolar Concentration; D010449:Peptide Mapping; D010766:Phosphorylation; D011189:Potassium Chloride / Q000494:pharmacology; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000254:growth & development*; D015398:Signal Transduction / Q000502:physiology; D012965:Sodium Chloride / Q000494:pharmacology*; D013912:Threonine / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D026341:Minichromosome Maintenance 1 Protein; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D013912:Threonine; D012965:Sodium Chloride; D011189:Potassium Chloride; D000135:Acid Phosphatase; D018021:Lithium Chloride