Functional analysis of the Arabidopsis thaliana mismatch repair gene MSH2.
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.
Adé
J
J
Département de phytologie, Pavillon C.-E. Marchand, Université Laval, Ste-Foy, QC, Canada.
Haffani
Y
Y
Beizile
F J
FJ
D000251:Adenosine Triphosphatases; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D001426:Bacterial Proteins / Q000235:genetics; D020137:Base Pair Mismatch; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004260:DNA Repair; D004268:DNA-Binding Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D015151:Immunoblotting; D051719:MutS DNA Mismatch-Binding Protein; D051718:MutS Homolog 2 Protein; D009154:Mutation; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D013696:Temperature
D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA; D000251:Adenosine Triphosphatases; C108984:MSH2 protein, Arabidopsis; D051719:MutS DNA Mismatch-Binding Protein; D051718:MutS Homolog 2 Protein; C497175:MutS protein, E coli; D012293:Rifampin