Bypass of a protein barrier by a replicative DNA helicase

Nature. 2012 Dec 13;492(7428):205-9. doi: 10.1038/nature11730. Epub 2012 Nov 28.

Abstract

Replicative DNA helicases generally unwind DNA as a single hexamer that encircles and translocates along one strand of the duplex while excluding the complementary strand (known as steric exclusion). By contrast, large T antigen, the replicative DNA helicase of the simian virus 40 (SV40), is reported to function as a pair of stacked hexamers that pumps double-stranded DNA through its central channel while laterally extruding single-stranded DNA. Here we use single-molecule and ensemble assays to show that large T antigen assembled on the SV40 origin unwinds DNA efficiently as a single hexamer that translocates on single-stranded DNA in the 3'-to-5' direction. Unexpectedly, large T antigen unwinds DNA past a DNA-protein crosslink on the translocation strand, suggesting that the large T antigen ring can open to bypass bulky adducts. Together, our data underscore the profound conservation among replicative helicase mechanisms, and reveal a new level of plasticity in the interactions of replicative helicases with DNA damage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral, Tumor / metabolism
  • DNA Helicases / metabolism*
  • DNA Replication
  • DNA, Single-Stranded / metabolism
  • DNA, Viral / metabolism
  • Replication Origin / physiology
  • Simian virus 40 / enzymology*
  • Viral Proteins / metabolism

Substances

  • Antigens, Viral, Tumor
  • DNA, Single-Stranded
  • DNA, Viral
  • Viral Proteins
  • DNA Helicases