The RAG1 homeodomain recruits HMG1 and HMG2 to facilitate recombination signal sequence binding and to enhance the intrinsic DNA-bending activity of RAG1-RAG2

Mol Cell Biol. 1999 Oct;19(10):6532-42. doi: 10.1128/MCB.19.10.6532.

Abstract

V(D)J recombination is initiated by the specific binding of the RAG1-RAG2 (RAG1/2) complex to the heptamer-nonamer recombination signal sequences (RSS). Several steps of the V(D)J recombination reaction can be reconstituted in vitro with only RAG1/2 plus the high-mobility-group protein HMG1 or HMG2. Here we show that the RAG1 homeodomain directly interacts with both HMG boxes of HMG1 and HMG2 (HMG1,2). This interaction facilitates the binding of RAG1/2 to the RSS, mainly by promoting high-affinity binding to the nonamer motif. Using circular-permutation assays, we found that the RAG1/2 complex bends the RSS DNA between the heptamer and nonamer motifs. HMG1,2 significantly enhance the binding and bending of the 23RSS but are not essential for the formation of a bent DNA intermediate on the 12RSS. A transient increase of HMG1,2 concentration in transfected cells increases the production of the final V(D)J recombinants in vivo.

Publication types

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

MeSH terms

  • Binding Sites
  • DNA-Binding Proteins / metabolism
  • High Mobility Group Proteins / metabolism*
  • Homeodomain Proteins / metabolism*
  • Nucleic Acid Conformation*
  • Protein Binding
  • Receptors, Antigen / genetics*
  • Recombination, Genetic*

Substances

  • DNA-Binding Proteins
  • High Mobility Group Proteins
  • Homeodomain Proteins
  • Receptors, Antigen
  • V(D)J recombination activating protein 2
  • RAG-1 protein