Werner syndrome protein interacts with human flap endonuclease 1 and stimulates its cleavage activity

• Published in: The EMBO journal Vol. 20; no. 20; pp. 5791 - 5801
• Main Authors: Brosh Jr, Robert M., von Kobbe, Cayetano, Sommers, Joshua A., Karmakar, Parimal, Opresko, Patricia L., Piotrowski, Jason, Dianova, Irina, Dianov, Grigory L., Bohr, Vilhelm A.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 15.10.2001; Blackwell Publishing Ltd; Oxford University Press
• Subjects: Adenosine Triphosphatases - physiology; Amino acids; Catalysis; deoxyribonuclease; Deoxyribonucleic acid; DNA; DNA - metabolism; DNA Helicases - chemistry; DNA Helicases - physiology; DNA-Binding Proteins - physiology; Endodeoxyribonucleases - chemistry; Endodeoxyribonucleases - metabolism; Enzyme Activation; Exodeoxyribonucleases; Exonucleases - physiology; FEN-1 protein; Flap endonuclease 1; Flap Endonucleases; genomic instability; HeLa Cells; helicase; Humans; Macromolecular Substances; Peptide Fragments - metabolism; Proliferating Cell Nuclear Antigen - physiology; Protein Structure, Tertiary; Recombinant Fusion Proteins - metabolism; RecQ Helicases; replication; Replication Protein A; Werner syndrome; Werner Syndrome - genetics; Werner Syndrome Helicase; WRN protein
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/20.20.5791

Werner syndrome (WS) is a human premature aging disorder characterized by chromosomal instability. The cellular defects of WS presumably reflect compromised or aberrant function of a DNA metabolic pathway that under normal circumstances confers stability to the genome. We report a novel interaction of the WRN gene product with the human 5′ flap endonuclease/5′–3′ exonuclease (FEN‐1), a DNA structure‐specific nuclease implicated in DNA replication, recombination and repair. WS protein (WRN) dramatically stimulates the rate of FEN‐1 cleavage of a 5′ flap DNA substrate. The WRN–FEN‐1 functional interaction is independent of WRN catalytic function and mediated by a 144 amino acid domain of WRN that shares homology with RecQ DNA helicases. A physical interaction between WRN and FEN‐1 is demonstrated by their co‐immunoprecipitation from HeLa cell lysate and affinity pull‐down experiments using a recombinant C‐terminal fragment of WRN. The underlying defect of WS is discussed in light of the evidence for the interaction between WRN and FEN‐1.