Isolation and Characterization of Two Saccharomyces cerevisiae Genes Encoding Homologs of the Bacterial HexA and MutS Mismatch Repair Proteins

INVESTIGATIONS

Isolation and Characterization of Two Saccharomyces cerevisiae Genes Encoding Homologs of the Bacterial HexA and MutS Mismatch Repair Proteins

R. A. Reenan and R. D. Kolodner
Division of Cellular and Molecular Biology, The Dana-Farber Cancer Institute, Boston, Massachusetts 02115, and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115

Homologs of the Escherichia coli (mutL, S and uvrD) and Streptococcus pneumoniae (hexA, B) genes involved in mismatch repair are known in several distantly related organisms. Degenerate oligonucleotide primers based on conserved regions of E. coli MutS protein and its homologs from Salmonella typhimurium, S. pneumoniae and human were used in the polymerase chain reaction (PCR) to amplify and clone mutS/hexA homologs from Saccharomyces cerevisiae. Two DNA sequences were amplified whose deduced amino acid sequences both shared a high degree of homology with MutS. These sequences were then used to clone the full-length genes from a yeast genomic library. Sequence analysis of the two MSH genes (MSH = mutS homolog), MSH1 and MSH2, revealed open reading frames of 2877 bp and 2898 bp. The deduced amino acid sequences predict polypeptides of 109.3 kD and 109.1 kD, respectively. The overall amino acid sequence identity with the E. coli MutS protein is 28.6% for MSH1 and 25.2% for MSH2. Features previously found to be shared by MutS homologs, such as the nucleotide binding site and the helix-turn-helix DNA binding motif as well as other highly conserved regions whose function remain unknown, were also found in the two yeast homologs. Evidence presented in this and a companion study suggest that MSH1 is involved in repair of mitochondrial DNA and that MSH2 is involved in nuclear DNA repair.

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				X. M. Zuo, G. D. Clark-Walker, and X. J. Chen The Mitochondrial Nucleoid Protein, Mgm101p, of Saccharomyces cerevisiae Is Involved in the Maintenance of {rho}+ and ori/rep-Devoid Petite Genomes but Is Not Required for Hypersuppressive {rho}- mtDNA Genetics, April 1, 2002; 160(4): 1389 - 1400. [Abstract] [Full Text] [PDF]

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				D. M. Wloch, K. Szafraniec, R. H. Borts, and R. Korona Direct Estimate of the Mutation Rate and the Distribution of Fitness Effects in the Yeast Saccharomyces cerevisiae Genetics, October 1, 2001; 159(2): 441 - 452. [Abstract] [Full Text] [PDF]

				C. Tornier, S. Bessone, I. Varlet, C. Rudolph, M. Darmon, and O. Fleck Requirement for Msh6, but Not for Swi4 (Msh3), in Msh2-Dependent Repair of Base-Base Mismatches and Mononucleotide Loops in Schizosaccharomyces pombe Genetics, May 1, 2001; 158(1): 65 - 75. [Abstract] [Full Text]

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				V. Contamine and M. Picard Maintenance and Integrity of the Mitochondrial Genome: a Plethora of Nuclear Genes in the Budding Yeast Microbiol. Mol. Biol. Rev., June 1, 2000; 64(2): 281 - 315. [Abstract] [Full Text] [PDF]

				K. M. Culligan, G. Meyer-Gauen, J. Lyons-Weiler, and J. B. Hays Evolutionary origin, diversification and specialization of eukaryotic MutS homolog mismatch repair proteins Nucleic Acids Res., January 15, 2000; 28(2): 463 - 471. [Abstract] [Full Text] [PDF]

				A. Nicholson, M. Hendrix, S. Jinks-Robertson, and G. F. Crouse Regulation of Mitotic Homeologous Recombination in Yeast: Functions of Mismatch Repair and Nucleotide Excision Repair Genes Genetics, January 1, 2000; 154(1): 133 - 146. [Abstract] [Full Text]

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				S. Guerrette, T. Wilson, S. Gradia, and R. Fishel Interactions of Human hMSH2 with hMSH3 and hMSH2 with hMSH6: Examination of Mutations Found in Hereditary Nonpolyposis Colorectal Cancer Mol. Cell. Biol., November 1, 1998; 18(11): 6616 - 6623. [Abstract] [Full Text]

				J. H. Cho, S. J. Ha, L. R. Kao, T. L. Megraw, and C.-B. Chae A Novel DNA-Binding Protein Bound to the Mitochondrial Inner Membrane Restores the Null Mutation of Mitochondrial Histone Abf2p in Saccharomyces cerevisiae Mol. Cell. Biol., October 1, 1998; 18(10): 5712 - 5723. [Abstract] [Full Text]

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				R. Fishel Mismatch repair, molecular switches, and signal transduction Genes & Dev., July 15, 1998; 12(14): 2096 - 2101. [Full Text]

				I. V. Fedorova, L. M. Gracheva, S. V. Kovaltzova, T. A. Evstuhina, S. Yu. Alekseev, and V. G. Korolev The Yeast HSM3 Gene Acts in One of the Mismatch Repair Pathways Genetics, March 1, 1998; 148(3): 963 - 973. [Abstract] [Full Text] [PDF]

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