The yeast RME1 gene encodes a putative zinc finger protein that is directly repressed by a1-alpha 2.
In the yeast Saccharomyces cerevisiae, a/alpha cells can enter meiosis whereas a and alpha cells cannot. The a/alpha cell type is determined by presence of a repressor, a1-alpha 2. Previous studies indicate that a/alpha cells lack an inhibitor of meiosis, the RME1 gene product, and that a and alpha cells express RME1. We report here the sequence of RME1 and functional analysis of its regulatory and coding regions. The 5'-region of RME1 includes a sequence resembling a1-alpha 2 repression sites. Deletion of this site at RME1 relieves repression by a1-alpha 2, and insertion of the site into a heterologous regulatory region (CYC1) confers weak repression in a/alpha cells. These observations indicate that RME1 is directly repressed by a1-alpha 2. The RME1 product has three regions that resemble C2H2 zinc fingers, which are characteristic of a class of nucleic-acid-binding proteins. Substitution of serine for cysteine in each of the putative fingers abolishes RME1 function; serine substitutions in the second and third putative fingers do not affect RME1 stability. These findings indicate that at least two putative zinc fingers are critical for RME1 structure or activity. Therefore RME1, which is formally a negative regulator of the meiotic gene IME1, may act directly as a repressor.
Covitz
P A
PA
Institute of Cancer Research, Columbia University, New York, New York 10032.
Herskowitz
I
I
Mitchell
A P
AP
D000595:Amino Acid Sequence; D001483:Base Sequence; D016384:Consensus Sequence; D003545:Cysteine / Q000737:chemistry; D004252:DNA Mutational Analysis; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D015151:Immunoblotting; D008540:Meiosis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D012689:Sequence Homology, Nucleic Acid; D012694:Serine / Q000737:chemistry; D016335:Zinc Fingers / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D005656:Fungal Proteins; C071594:RME1 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D012694:Serine; D001616:beta-Galactosidase; D003545:Cysteine