RAG1-mediated ubiquitylation of histone H3 is required for chromosomal V(D)J recombination

Cell Res. 2015 Feb;25(2):181-92. doi: 10.1038/cr.2015.1. Epub 2015 Jan 9.

Abstract

RAG1 and RAG2 proteins are key components in V(D)J recombination. The core region of RAG1 is capable of catalyzing the recombination reaction; however, the biological function of non-core RAG1 remains largely unknown. Here, we show that in a murine-model carrying the RAG1 ring-finger conserved cysteine residue mutation (C325Y), V(D)J recombination was abrogated at the cleavage step, and this effect was accompanied by decreased mono-ubiquitylation of histone H3. Further analyses suggest that un-ubiquitylated histone H3 restrains RAG1 to the chromatin by interacting with the N-terminal 218 amino acids of RAG1. Our data provide evidence for a model in which ubiquitylation of histone H3 mediated by the ring-finger domain of RAG1 triggers the release of RAG1, thus allowing its transition into the cleavage phase. Collectively, our findings reveal that the non-core region of RAG1 facilitates chromosomal V(D)J recombination in a ubiquitylation-dependent pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • Chromosomes / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism
  • Female
  • HEK293 Cells
  • Histones / metabolism*
  • Homeodomain Proteins / chemistry
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Protein Structure, Tertiary
  • Receptors, Antigen, T-Cell, alpha-beta / metabolism
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • Transfection
  • Ubiquitination
  • V(D)J Recombination / physiology*

Substances

  • DNA-Binding Proteins
  • Histones
  • Homeodomain Proteins
  • Rag2 protein, mouse
  • Receptors, Antigen, T-Cell, alpha-beta
  • RAG-1 protein