Werner protein stimulates topoisomerase I DNA relaxation activity.
Werner syndrome (WS) is a human premature aging disorder characterized by the early onset of age-related clinical features and an elevated incidence of cancer. The Werner protein (WRN) belongs to the RecQ family of DNA helicases and is required for the maintenance of genomic stability in human cells. Potential cooperation between RecQ helicases and topoisomerases in many aspects of DNA metabolism, such as the progression of replication forks, transcription, recombination, and repair, has been reported. Here, we show a physical and functional interaction between WRN and topoisomerase I (topo I). WRN colocalizes and interacts directly with topo I. WRN stimulates the ability of topo I to relax negatively supercoiled DNA and specifically stimulates the religation step of the relaxation reaction. Moreover, cell extracts from WS fibroblasts exhibit a decrease in the relaxation activity of negatively supercoiled DNA. We have identified two regions of WRN that mediate functional interaction with topo I, and they are located at the NH(2) and COOH termini of the WRN protein. In a reciprocal functional interaction, topo I inhibits the ATPase activity of WRN. Our data provide new insight into the interrelationship between RecQ helicases and topoisomerases in the maintenance of genomic integrity and prevention of tumorigenesis.
Laine
Jean-Philippe
JP
Laboratory of Molecular Gerontology, National Institute on Aging/NIH, 5600 Nathan Shock Drive, Baltimore, MD 21224, USA.
Opresko
Patricia L
PL
Indig
Fred E
FE
Harrigan
Jeanine A
JA
von Kobbe
Cayetano
C
Bohr
Vilhelm A
VA
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D002166:Camptothecin / Q000494:pharmacology; D002478:Cells, Cultured; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004278:DNA, Superhelical / Q000378:metabolism; D005090:Exodeoxyribonucleases; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D010449:Peptide Mapping; D017434:Protein Structure, Tertiary; D053484:RecQ Helicases; D014898:Werner Syndrome / Q000201:enzymology / Q000473:pathology; D000071657:Werner Syndrome Helicase
D004278:DNA, Superhelical; D005090:Exodeoxyribonucleases; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin