Mutations affecting a putative MutLalpha endonuclease motif impact multiple mismatch repair functions.
Mutations in DNA mismatch repair (MMR) lead to increased mutation rates and higher recombination between similar, but not identical sequences, as well as resistance to certain DNA methylating agents. Recently, a component of human MMR machinery, MutLalpha, has been shown to display a latent endonuclease activity. The endonuclease active site appears to include a conserved motif, DQHA(X)(2)E(X)(4)E, within the COOH-terminus of human PMS2. Substitution of the glutamic acid residue (E705) abolished the endonuclease activity and mismatch-dependent excision in vitro. Previously, we showed that the PMS2-E705K mutation and the corresponding mutation in Saccharomyces cerevisiae were both recessive loss of function alleles for mutation avoidance in vivo. Here, we show that mutations impacting this endonuclease motif also significantly affect MMR-dependent suppression of homeologous recombination in yeast and responses to S(n)1-type methylating agents in both yeast and mammalian cells. Thus, our in vivo results suggest that the endonuclease activity of MutLalpha is important not only in MMR-dependent mutation avoidance but also for recombination and damage response functions.
Erdeniz
Naz
N
Department of Molecular and Medical Genetics, Oregon Health & Science University L103, 3181 SW Sam Jackson Park Road, Portland, OR 97239-3098, USA.
Nguyen
Megan
M
Deschênes
Suzanne M
SM
Liskay
R Michael
RM
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D004249:DNA Damage; D017931:DNA Primers; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000235:genetics*; D006801:Humans; D051379:Mice; D000070956:MutL Proteins; D009154:Mutation; D011995:Recombination, Genetic
D017931:DNA Primers; C500929:MutLalpha protein, human; D000070956:MutL Proteins; D045643:DNA Repair Enzymes