RPA accumulation during class switch recombination represents 5'-3' DNA-end resection during the S-G2/M phase of the cell cycle.

Abstract

Activation-induced cytidine deaminase (AID) promotes chromosomal translocations by inducing DNA double-strand breaks (DSBs) at immunoglobulin (Ig) genes and oncogenes in the G1 phase. RPA is a single-stranded DNA (ssDNA)-binding protein that associates with resected DSBs in the S phase and facilitates the assembly of factors involved in homologous repair (HR), such as Rad51. Notably, RPA deposition also marks sites of AID-mediated damage, but its role in Ig gene recombination remains unclear. Here, we demonstrate that RPA associates asymmetrically with resected ssDNA in response to lesions created by AID, recombination-activating genes (RAG), or other nucleases. Small amounts of RPA are deposited at AID targets in G1 in an ATM-dependent manner. In contrast, recruitment in the S-G2/M phase is extensive, ATM independent, and associated with Rad51 accumulation. In the S-G2/M phase, RPA increases in nonhomologous-end-joining-deficient lymphocytes, where there is more extensive DNA-end resection. Thus, most RPA recruitment during class switch recombination represents salvage of unrepaired breaks by homology-based pathways during the S-G2/M phase of the cell cycle.

• Authors: Arito Yamane, Davide F Robbiani, Wolfgang Resch, Anne Bothmer, Hirotaka Nakahashi, Thiago Oliveira, Philipp C Rommel, Eric J Brown, Andre Nussenzweig, Michel C Nussenzweig, Rafael Casellas
• Journal: Cell reports (Cell Rep) Vol. 3 Issue 1 Pg. 138-47 (Jan 31 2013) ISSN: 2211-1247 [Electronic] United States
• PMID: 23291097 (Publication Type: Journal Article, Research Support, N.I.H., Intramural)
• Copyright: Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Cell Cycle Proteins
• DNA, Single-Stranded
• DNA-Binding Proteins
• Histones
• Replication Protein A
• Tumor Suppressor Proteins
• gamma-H2AX protein, mouse
• Ataxia Telangiectasia Mutated Proteins
• Atm protein, mouse
• Protein Serine-Threonine Kinases
• Rad51 Recombinase
• Cytidine Deaminase

Topics

• Animals
• Ataxia Telangiectasia Mutated Proteins
• Cell Cycle Proteins (metabolism)
• Chromatin Immunoprecipitation
• Cytidine Deaminase (metabolism)
• DNA Breaks
• DNA Breaks, Double-Stranded
• DNA, Single-Stranded (metabolism)
• DNA-Binding Proteins (metabolism)
• G2 Phase (genetics)
• Genetic Loci
• Histones (metabolism)
• Immunoglobulin Class Switching (genetics)
• Mice
• Models, Biological
• Protein Binding (genetics)
• Protein Serine-Threonine Kinases (metabolism)
• Rad51 Recombinase (metabolism)
• Recombination, Genetic
• Replication Protein A (metabolism)
• S Phase (genetics)
• Tumor Suppressor Proteins (metabolism)