Functional analysis of the Arabidopsis thaliana mismatch repair gene MSH2.

J Adé, Y Haffani, F J Beizile

Journal: Genome 2002;44(4):651-7

PMID: 11550901

Abstract

The Arabidopsis thaliana MSH2 (AtMSH2) gene encodes a protein that belongs to a family of highly conserved proteins (MutS homologues (MSH)) involved in DNA mismatch repair. Sequence analysis strongly suggests that this single copy gene is indeed a homologue of MSH2, a gene known to play a central role in eukaryotic mismatch repair. In this report, we show that the AtMSH2 protein has functional attributes characteristic of previously described mismatch repair proteins. First, over-expression of this protein in Escherichia coli leads to a mutator phenotype similar to that reported previously for known functional homologues. Second, gel retardation assays revealed that the AtMSH2 protein has a 10-fold greater affinity for DNA containing a single pair of mismatched nucleotides versus perfectly matched DNA. These results provide experimental evidence that AtMSH2 is indeed a functional homologue of MutS.

Address: Département de phytologie, Pavillon C.-E. Marchand, Université Laval, Ste-Foy, QC, Canada.

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MeSH Terms: Adenosine Triphosphatases, Arabidopsis, Arabidopsis Proteins, Bacterial Proteins, Base Pair Mismatch, Blotting, Western, Cloning, Molecular, DNA, DNA Repair, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Enzyme Inhibitors, Escherichia coli, Escherichia coli Proteins, Immunoblotting, MutS DNA Mismatch-Binding Protein, MutS Homolog 2 Protein, Mutation, Phenotype, Plant Proteins, Plasmids, Protein Binding, Recombinant Fusion Proteins, Rifampin, Temperature