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Werner syndrome helicase contains a 5'-->3' exonuclease activity that digests DNA and RNA strands in DNA/DNA and RNA/DNA duplexes dependent on unwinding.

Abstract
We show that WRN helicase contains a unique 5'-->3' exonuclease activity in the N-terminal region. Adeletion mutant lacking 231 N-terminal amino acid residues, made in a baculovirus system, did nothave this activity, while it showed ATPase and DNA helicase activities. This exonuclease activity was co-precipitated with the helicase activity using monoclonal antibodies specific to WRN helicase, indicating that it is an integral component with WRN helicase. The exonuclease in WRN helicase does not digest free single-stranded DNA or RNA, but it digests a strand in the duplex DNA or an RNA strand in a RNA/DNA heteroduplex in a 5'-->3' direction dependent on duplex unwinding. The digestion products were identified as 5'-mononucleotides. Our data show that WRN helicase needs a single-stranded 3' overhang region for efficient binding and unwinding of duplex molecules, while blunt-ended or 5' overhang duplex molecules were hardly unwound. These findings suggest that the WRN helicase and integral 5'-->3' exonuclease activities are involved in preventing a hyper-recombination by resolving entangled structures of DNA and RNA/DNA heteroduplexes that may be generated during rep-lication, repair and/or transcription.
AuthorsN Suzuki, M Shiratori, M Goto, Y Furuichi
JournalNucleic acids research (Nucleic Acids Res) Vol. 27 Issue 11 Pg. 2361-8 (Jun 01 1999) ISSN: 0305-1048 [Print] England
PMID10325426 (Publication Type: Journal Article)
Chemical References
  • Antibodies, Monoclonal
  • Nucleic Acid Heteroduplexes
  • RNA
  • DNA
  • Exodeoxyribonucleases
  • Exodeoxyribonuclease V
  • DNA Helicases
  • RecQ Helicases
  • WRN protein, human
  • Werner Syndrome Helicase
Topics
  • Animals
  • Antibodies, Monoclonal
  • Cell Line
  • DNA (metabolism)
  • DNA Helicases (genetics, metabolism)
  • Exodeoxyribonuclease V
  • Exodeoxyribonucleases (metabolism)
  • Humans
  • Mutagenesis, Site-Directed
  • Nucleic Acid Heteroduplexes (metabolism)
  • Precipitin Tests
  • RNA (metabolism)
  • RecQ Helicases
  • Spodoptera (cytology)
  • Werner Syndrome (enzymology)
  • Werner Syndrome Helicase

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