3'-processing and strand transfer catalysed by retroviral integrase in crystallo

EMBO J. 2012 Jun 29;31(13):3020-8. doi: 10.1038/emboj.2012.118. Epub 2012 May 11.

Abstract

Retroviral integrase (IN) is responsible for two consecutive reactions, which lead to insertion of a viral DNA copy into a host cell chromosome. Initially, the enzyme removes di- or trinucleotides from viral DNA ends to expose 3'-hydroxyls attached to the invariant CA dinucleotides (3'-processing reaction). Second, it inserts the processed 3'-viral DNA ends into host chromosomal DNA (strand transfer). Herein, we report a crystal structure of prototype foamy virus IN bound to viral DNA prior to 3'-processing. Furthermore, taking advantage of its dependence on divalent metal ion cofactors, we were able to freeze trap the viral enzyme in its ground states containing all the components necessary for 3'-processing or strand transfer. Our results shed light on the mechanics of retroviral DNA integration and explain why HIV IN strand transfer inhibitors are ineffective against the 3'-processing step of integration. The ground state structures moreover highlight a striking substrate mimicry utilized by the inhibitors in their binding to the IN active site and suggest ways to improve upon this clinically relevant class of small molecules.

Publication types

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

MeSH terms

  • Catalysis
  • Catalytic Domain
  • Cations, Divalent / chemistry
  • DNA, Viral / metabolism
  • HIV / enzymology
  • Integrases / chemistry*
  • Spumavirus / enzymology*
  • Virus Integration
  • X-Ray Diffraction

Substances

  • Cations, Divalent
  • DNA, Viral
  • Integrases