Document Detail


Genetic and biochemical analysis of Msh2p-Msh6p: role of ATP hydrolysis and Msh2p-Msh6p subunit interactions in mismatch base pair recognition.
MedLine Citation:
PMID:  9111312     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
E Alani; T Sokolsky; B Studamire; J J Miret; R S Lahue
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular and cellular biology     Volume:  17     ISSN:  0270-7306     ISO Abbreviation:  Mol. Cell. Biol.     Publication Date:  1997 May 
Date Detail:
Created Date:  1997-05-15     Completed Date:  1997-05-15     Revised Date:  2014-04-08    
Medline Journal Info:
Nlm Unique ID:  8109087     Medline TA:  Mol Cell Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2436-47     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphatases / metabolism
Adenosine Triphosphate / metabolism*
Alleles
Amino Acid Sequence
Bacteriophage lambda
Binding Sites
DNA Repair*
DNA-Binding Proteins / metabolism*
Fungal Proteins / metabolism*
Helix-Turn-Helix Motifs
Humans
Hydrolysis
Models, Molecular
Molecular Sequence Data
MutS Homolog 2 Protein
Mutagenesis, Site-Directed
Nucleic Acid Heteroduplexes / metabolism*
Phenotype
Proto-Oncogene Proteins / metabolism*
Repressor Proteins / metabolism
Saccharomyces cerevisiae Proteins*
Sequence Alignment
Viral Proteins
Viral Regulatory and Accessory Proteins
Grant Support
ID/Acronym/Agency:
GM53085/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/DNA-Binding Proteins; 0/Fungal Proteins; 0/MSH6 protein, S cerevisiae; 0/Nucleic Acid Heteroduplexes; 0/Proto-Oncogene Proteins; 0/Repressor Proteins; 0/Saccharomyces cerevisiae Proteins; 0/Viral Proteins; 0/Viral Regulatory and Accessory Proteins; 0/phage repressor proteins; 8L70Q75FXE/Adenosine Triphosphate; EC 3.6.1.-/Adenosine Triphosphatases; EC 3.6.1.3/MSH2 protein, S cerevisiae; EC 3.6.1.3/MSH2 protein, human; EC 3.6.1.3/MutS Homolog 2 Protein
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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