Interaction of HIV-1 integrase with DNA repair protein hRad18

J Biol Chem. 2002 Jul 26;277(30):27489-93. doi: 10.1074/jbc.M203061200. Epub 2002 May 16.

Abstract

We have previously shown that human immunodeficiency virus-1 (HIV-1) integrase is an unstable protein and a substrate for the N-end rule degradation pathway. This degradation pathway shares its ubiquitin-conjugating enzyme, Rad6, with the post-replication/translesion DNA repair pathway. Because DNA repair is thought to play an essential role in HIV-1 integration, we investigated whether other molecules of this DNA repair pathway could interact with integrase. We observed that co-expression of human Rad18 induced the accumulation of an otherwise unstable form of HIV-1 integrase. This accumulation occurred even though hRAD18 possesses a RING finger domain, a structure that is generally associated with E3 ubiquitin ligase function and protein degradation. Evidence for an interaction between integrase and hRad18 was obtained through reciprocal co-immunoprecipitation. Moreover we found that a 162-residue region of hRad18 (amino acids 65-226) was sufficient for both integrase stabilization and interaction. Finally, we observed that HIV-1 integrase co-localized with hRad18 in nuclear structures in a subpopulation of co-transfected cells. Taken together, these findings identify hRad18 as a novel interacting partner of HIV-1 integrase and suggest a role for post-replication/translesion DNA repair in the retroviral integration process.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Blotting, Western
  • Cell Line
  • DNA Repair*
  • DNA-Binding Proteins / metabolism*
  • Gene Deletion
  • HIV Integrase / metabolism*
  • Humans
  • Microscopy, Fluorescence
  • Mutation
  • Plasmids / metabolism
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Retroviridae / genetics
  • Transfection
  • Ubiquitin-Protein Ligases

Substances

  • DNA-Binding Proteins
  • RAD18 protein, human
  • Ubiquitin-Protein Ligases
  • HIV Integrase