Physical and functional interactions between Werner syndrome helicase and mismatch-repair initiation factors.
Werner syndrome (WS) is a severe recessive disorder characterized by premature aging, cancer predisposition and genomic instability. The gene mutated in WS encodes a bi-functional enzyme called WRN that acts as a RecQ-type DNA helicase and a 3'-5' exonuclease, but its exact role in DNA metabolism is poorly understood. Here we show that WRN physically interacts with the MSH2/MSH6 (MutSalpha), MSH2/MSH3 (MutSbeta) and MLH1/PMS2 (MutLalpha) heterodimers that are involved in the initiation of mismatch repair (MMR) and the rejection of homeologous recombination. MutSalpha and MutSbeta can strongly stimulate the helicase activity of WRN specifically on forked DNA structures with a 3'-single-stranded arm. The stimulatory effect of MutSalpha on WRN-mediated unwinding is enhanced by a G/T mismatch in the DNA duplex ahead of the fork. The MutLalpha protein known to bind to the MutS alpha-heteroduplex complexes has no effect on WRN-mediated DNA unwinding stimulated by MutSalpha, nor does it affect DNA unwinding by WRN alone. Our data are consistent with results of genetic experiments in yeast suggesting that MMR factors act in conjunction with a RecQ-type helicase to reject recombination between divergent sequences.
Saydam
Nurten
N
Institute of Molecular Cancer Research of the University of Zurich, Switzerland.
Kanagaraj
Radhakrishnan
R
Dietschy
Tobias
T
Garcia
Patrick L
PL
Peña-Diaz
Javier
J
Shevelev
Igor
I
Stagljar
Igor
I
Janscak
Pavel
P
D020137:Base Pair Mismatch; D001665:Binding Sites; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005090:Exodeoxyribonucleases; D006801:Humans; D000070956:MutL Proteins; D051718:MutS Homolog 2 Protein / Q000378:metabolism; D000074081:MutS Homolog 3 Protein; D017434:Protein Structure, Tertiary; D053484:RecQ Helicases / Q000737:chemistry / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D000071657:Werner Syndrome Helicase
D004268:DNA-Binding Proteins; C094443:G-T mismatch-binding protein; C078444:MSH3 protein, human; C500929:MutLalpha protein, human; D000074081:MutS Homolog 3 Protein; D004247:DNA; D005090:Exodeoxyribonucleases; C497172:MSH2 protein, human; D000070956:MutL Proteins; D051718:MutS Homolog 2 Protein; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase; D045643:DNA Repair Enzymes
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