A role of the C-terminal region adjacent to the zinc-fingers in the DNA binding ability of Rme1p, a regulator of meiosis in S. cerevisiae.
Rme1p is a zinc-finger protein and has a pivotal role in control of meiosis in Saccharomyces cerevisiae. The DNA binding domain of Rme1p consists of three zinc-finger segments and the C-terminal 16 amino acid residues (called C-TR). To examine the role of C-TR, a series of mutant Rme1p fused with maltose binding protein (MBP) were constructed, purified, and characterized, in terms of the DNA binding ability. The basic amino acid residues R287 and K290, and the hydrophobic residues F288, L292, 1295, and L296 play an important role for DNA binding, suggesting that the C-TR forms an amphipathic alpha-helix. Also, it was shown that the mutations in the basic amino acid residues abolish the repression and inhibition of spore formation by Rme1p in vivo. Hence, the C-TR is important for in vivo function of Rme1p.
Shimizu
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M
Department of Chemistry, Meisei University, Hino, Tokyo, Japan.
Murase
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A
Shindo
H
H
Mitchell
A P
AP
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D008540:Meiosis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012097:Repressor Proteins; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D016335:Zinc Fingers
D005656:Fungal Proteins; D009838:Oligodeoxyribonucleotides; C071594:RME1 protein, S cerevisiae; D011994:Recombinant Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA