Title

Deletion mutation analysis of the mutS gene in Escherichia coli

UMMS Affiliation

Department of Pharmacology and Molecular Toxicology

Date

2-20-1999

Document Type

Article

Subjects

*Adenosine Triphosphatases; Amino Acid Sequence; Bacterial Proteins; Base Sequence; DNA Repair; *DNA-Binding Proteins; Escherichia coli; *Escherichia coli Proteins; Molecular Sequence Data; MutS DNA Mismatch-Binding Protein; *Mutation; Nucleic Acid Heteroduplexes; Protein Binding; *Sequence Deletion

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

The MutS protein is part of the dam-directed MutHLS mismatch repair pathway in Escherichia coli. We have constructed deletion derivatives in the mutS gene, which retain the P-loop coding region for ATP binding. The mutant proteins were assayed for ATP hydrolysis, heteroduplex DNA binding, heterodimer MutS formation, and the ability to interact with MutL. Dimerization was assayed by expressing His6-tagged wild-type and non-tagged deletion mutant proteins in the same cell and isolating the His6-tagged protein followed by MutS immunoblotting after SDS-polyacrylamide gel electrophoresis. MutS-MutL interaction was measured using the same technique except that the MutL protein carried the His6 tag. Our results indicate that DNA binding ability resides in the N-terminal end of MutS, and dimerization and MutL interactions are located in the C-terminal end. Given the extensive amino acid homology in the MutS family our results with E. coli should be applicable to MutS homologues in other prokaryotes and eukaryotes.

Rights and Permissions

Citation: J Biol Chem. 1999 Feb 26;274(9):5948-52.

Related Resources

Link to Article in PubMed

Journal Title

The Journal of biological chemistry

PubMed ID

10026220