This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Alani, E. Articles by Lahue, R. S. Search for Related Content PubMed PubMed Citation Articles by Alani, E. Articles by Lahue, R. S.

 Previous Article  |  Next Article 

Mol. Cell. Biol., May 1997, 2436-2447, Vol 17, No. 5
Copyright © 1997, American Society for Microbiology

Genetic and biochemical analysis of Msh2p-Msh6p: role of ATP hydrolysis and Msh2p-Msh6p subunit interactions in mismatch base pair recognition

E Alani, T Sokolsky, B Studamire, JJ Miret and RS Lahue
Section of Genetics and Development, Cornell University, Ithaca, New York 14853-2703, USA. eea3@cornell.edu

Recent studies have shown that Saccharomyces cerevisiae Msh2p and Msh6p form a complex that specifically binds to DNA containing base pair mismatches. In this study, we performed a genetic and biochemical analysis of the Msh2p-Msh6p complex by introducing point mutations in the ATP binding and putative helix-turn-helix domains of MSH2. The effects of these mutations were analyzed genetically by measuring mutation frequency and biochemically by measuring the stability, mismatch binding activity, and ATPase activity of msh2p (mutant msh2p)- Msh6p complexes. A mutation in the ATP binding domain of MSH2 did not affect the mismatch binding specificity of the msh2p-Msh6p complex; however, this mutation conferred a dominant negative phenotype when the mutant gene was overexpressed in a wild-type strain, and the mutant protein displayed biochemical defects consistent with defects in mismatch repair downstream of mismatch recognition. Helix-turn-helix domain mutant proteins displayed two different properties. One class of mutant proteins was defective in forming complexes with Msh6p and also failed to recognize base pair mismatches. A second class of mutant proteins displayed properties similar to those observed for the ATP binding domain mutant protein. Taken together, these data suggested that the proposed helix-turn-helix domain of Msh2p was unlikely to be involved in mismatch recognition. We propose that the MSH2 helix-turn- helix domain mediates changes in Msh2p-Msh6p interactions that are induced by ATP hydrolysis; the net result of these changes is a modulation of mismatch recognition.

This article has been cited by other articles:

• Gruver, A. M., Miller, K. A., Rajesh, C., Smiraldo, P. G., Kaliyaperumal, S., Balder, R., Stiles, K. M., Albala, J. S., Pittman, D. L. (2005). The ATPase motif in RAD51D is required for resistance to DNA interstrand crosslinking agents and interaction with RAD51C. Mutagenesis 20: 433-440 [Abstract] [Full Text]  
• Lin, D. P., Wang, Y., Scherer, S. J., Clark, A. B., Yang, K., Avdievich, E., Jin, B., Werling, U., Parris, T., Kurihara, N., Umar, A., Kucherlapati, R., Lipkin, M., Kunkel, T. A., Edelmann, W. (2004). An Msh2 Point Mutation Uncouples DNA Mismatch Repair and Apoptosis. Cancer Res. 64: 517-522 [Abstract] [Full Text]  
• Leonard, J. M., Bollmann, S. R., Hays, J. B. (2003). Reduction of Stability of Arabidopsis Genomic and Transgenic DNA-Repeat Sequences (Microsatellites) by Inactivation of AtMSH2 Mismatch-Repair Function. Plant Physiol. 133: 328-338 [Abstract] [Full Text]  
• Plotz, G., Raedle, J., Brieger, A., Trojan, J., Zeuzem, S. (2003). N-terminus of hMLH1 confers interaction of hMutL{alpha} and hMutL{beta} with hMutS{alpha}. Nucleic Acids Res 31: 3217-3226 [Abstract] [Full Text]  
• Pezza, R. J., Villarreal, M. A., Montich, G. G., Argarana, C. E. (2002). Vanadate inhibits the ATPase activity and DNA binding capability of bacterial MutS. A structural model for the vanadate-MutS interaction at the Walker A motif. Nucleic Acids Res 30: 4700-4708 [Abstract] [Full Text]  
• Alas, M. M. d. l., Los, G., Lin, X., Kurdi-Haidar, B., Manorek, G., Howell, S. B. (2002). Identification of Transdominant-Negative Genetic Suppressor Elements Derived from hMSH2 That Mediate Resistance to 6-Thioguanine. Mol. Pharmacol. 62: 1198-1206 [Abstract] [Full Text]  
• Hess, M. T., Gupta, R. D., Kolodner, R. D. (2002). Dominant Saccharomyces cerevisiae msh6 Mutations Cause Increased Mispair Binding and Decreased Dissociation from Mispairs by Msh2-Msh6 in the Presence of ATP. J. Biol. Chem. 277: 25545-25553 [Abstract] [Full Text]  
• Christmann, M., Tomicic, M. T., Kaina, B. (2002). Phosphorylation of mismatch repair proteins MSH2 and MSH6 affecting MutS{alpha} mismatch-binding activity. Nucleic Acids Res 30: 1959-1966 [Abstract] [Full Text]  
• Plotz, G., Raedle, J., Brieger, A., Trojan, J., Zeuzem, S. (2002). hMutS{alpha} forms an ATP-dependent complex with hMutL{alpha} and hMutL{beta} on DNA. Nucleic Acids Res 30: 711-718 [Abstract] [Full Text]  
• Drotschmann, K., Yang, W., Brownewell, F. E., Kool, E. T., Kunkel, T. A. (2001). Asymmetric Recognition of DNA Local Distortion. STRUCTURE-BASED FUNCTIONAL STUDIES OF EUKARYOTIC Msh2-Msh6. J. Biol. Chem. 276: 46225-46229 [Abstract] [Full Text]  
• Pang, Q., Christianson, T. A., Keeble, W., Diaz, J., Faulkner, G. R., Reifsteck, C., Olson, S., Bagby, G. C. (2001). The Fanconi anemia complementation group C gene product: structural evidence of multifunctionality. Blood 98: 1392-1401 [Abstract] [Full Text]  
• Tran, P. T., Simon, J. A., Liskay, R. M. (2001). Interactions of Exo1p with components of MutLalpha in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 10.1073/pnas.161175998v1 [Abstract] [Full Text]  
• Kelly, K. O., Dernburg, A. F., Stanfield, G. M., Villeneuve, A. M. (2000). Caenorhabditis elegans msh-5 Is Required for Both Normal and Radiation-Induced Meiotic Crossing Over but Not for Completion of Meiosis. Genetics 156: 617-630 [Abstract] [Full Text]  
• Tran, P. T., Liskay, R. M. (2000). Functional Studies on the Candidate ATPase Domains of Saccharomyces cerevisiae MutLalpha. Mol. Cell. Biol. 20: 6390-6398 [Abstract] [Full Text]  
• Sokolsky, T., Alani, E. (2000). EXO1 and MSH6 Are High-Copy Suppressors of Conditional Mutations in the MSH2 Mismatch Repair Gene of Saccharomyces cerevisiae. Genetics 155: 589-599 [Abstract] [Full Text]  
• Joshi, A., Sen, S., Rao, B. J. (2000). ATP-hydrolysis-dependent conformational switch modulates the stability of MutS-mismatch complexes. Nucleic Acids Res 28: 853-861 [Abstract] [Full Text]  
• Gradia, S., Acharya, S., Fishel, R. (2000). The Role of Mismatched Nucleotides in Activating the hMSH2-hMSH6 Molecular Switch. J. Biol. Chem. 275: 3922-3930 [Abstract] [Full Text]  
• Studamire, B., Price, G., Sugawara, N., Haber, J. E., Alani, E. (1999). Separation-of-Function Mutations in Saccharomyces cerevisiae MSH2 That Confer Mismatch Repair Defects but Do Not Affect Nonhomologous-Tail Removal during Recombination. Mol. Cell. Biol. 19: 7558-7567 [Abstract] [Full Text]  
• Zalevsky, J., MacQueen, A. J., Duffy, J. B., Kemphues, K. J., Villeneuve, A. M. (1999). Crossing Over During Caenorhabditis elegans Meiosis Requires a Conserved MutS-Based Pathway That Is Partially Dispensable in Budding Yeast. Genetics 153: 1271-1283 [Abstract] [Full Text]  
• Marsischky, G. T., Kolodner, R. D. (1999). Biochemical Characterization of the Interaction between the Saccharomyces cerevisiae MSH2-MSH6 Complex and Mispaired Bases in DNA. J. Biol. Chem. 274: 26668-26682 [Abstract] [Full Text]  
• Matton, N., Simonetti, J., Williams, K. (1999). Inefficient in vivo repair of mismatches at an oncogenic hotspot correlated with lack of binding by mismatch repair proteins and with phase of the cell cycle. Carcinogenesis 20: 1417-1424 [Abstract] [Full Text]  
• Wilson, T., Guerrette, S., Fishel, R. (1999). Dissociation of Mismatch Recognition and ATPase Activity by hMSH2-hMSH3. J. Biol. Chem. 274: 21659-21664 [Abstract] [Full Text]  
• Harfe, B. D., Jinks-Robertson, S. (1999). Removal of Frameshift Intermediates by Mismatch Repair Proteins in Saccharomyces cerevisiae. Mol. Cell. Biol. 19: 4766-4773 [Abstract] [Full Text]  
• Bowers, J., Sokolsky, T., Quach, T., Alani, E. (1999). A Mutation in the MSH6 Subunit of the Saccharomyces cerevisiae MSH2-MSH6 Complex Disrupts Mismatch Recognition. J. Biol. Chem. 274: 16115-16125 [Abstract] [Full Text]  
• Drotschmann, K., Clark, A. B., Tran, H. T., Resnick, M. A., Gordenin, D. A., Kunkel, T. A. (1999). Mutator phenotypes of yeast strains heterozygous for mutations in the MSH2 gene. Proc. Natl. Acad. Sci. USA 96: 2970-2975 [Abstract] [Full Text]  
• Wu, T.-H., Marinus, M. G. (1999). Deletion Mutation Analysis of the mutS Gene in Escherichia coli. J. Biol. Chem. 274: 5948-5952 [Abstract] [Full Text]  
• Studamire, B., Quach, T., Alani, E. (1998). Saccharomyces cerevisiae Msh2p and Msh6p ATPase Activities Are Both Required during Mismatch Repair. Mol. Cell. Biol. 18: 7590-7601 [Abstract] [Full Text]  
• Blackwell, L. J., Bjornson, K. P., Modrich, P. (1998). DNA-dependent Activation of the hMutSalpha ATPase. J. Biol. Chem. 273: 32049-32054 [Abstract] [Full Text]  
• Blackwell, L. J., Martik, D., Bjornson, K. P., Bjornson, E. S., Modrich, P. (1998). Nucleotide-promoted Release of hMutSalpha from Heteroduplex DNA Is Consistent with an ATP-dependent Translocation Mechanism. J. Biol. Chem. 273: 32055-32062 [Abstract] [Full Text]  
• Guerrette, S., Wilson, T., Gradia, S., Fishel, R. (1998). Interactions of Human hMSH2 with hMSH3 and hMSH2 with hMSH6: Examination of Mutations Found in Hereditary Nonpolyposis Colorectal Cancer. Mol. Cell. Biol. 18: 6616-6623 [Abstract] [Full Text]  
• Fishel, R. (1998). Mismatch repair, molecular switches, and signal transduction. Genes Dev. 12: 2096-2101 [Full Text]  
• De las Alas, M., de bruin, R. A. M., Eyck, L. T., Los, G., Howell, S. B. (1998). Prediction-based threading of the hMSH2 DNA mismatch repair protein. FASEB J. 12: 653-663 [Abstract] [Full Text]  
• Umar, A., Risinger, J. I., Glaab, W. E., Tindall, K. R., Barrett, J. C., Kunkel, T. A. (1998). Functional Overlap in Mismatch Repair by Human MSH3 and MSH6. Genetics 148: 1637-1646 [Abstract] [Full Text]  
• Pochart, P., Woltering, D., Hollingsworth, N. M. (1997). Conserved Properties between Functionally Distinct MutS Homologs in Yeast. J. Biol. Chem. 272: 30345-30349 [Abstract] [Full Text]  
• Degtyareva, N. P., Greenwell, P., Hofmann, E. R., Hengartner, M. O., Zhang, L., Culotti, J. G., Petes, T. D. (2002). Caenorhabditis elegans DNA mismatch repair gene msh-2 is required for microsatellite stability and maintenance of genome integrity. Proc. Natl. Acad. Sci. USA 99: 2158-2163 [Abstract] [Full Text]  
• Tran, P. T., Simon, J. A., Liskay, R. M. (2001). Interactions of Exo1p with components of MutLalpha in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 98: 9760-9765 [Abstract] [Full Text]