The ATPase activity of MLH1 is required to orchestrate DNA double-strand breaks and end processing during class switch recombination

Richard Chahwan, Johanna M M van Oers, Elena Avdievich, Chunfang Zhao, Winfried Edelmann, Matthew D. Scharff, Sergio Roa Roa

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Antibody diversification through somatic hypermutation (SHM) and class switch recombination (CSR) are similarly initiated in B cells with the generation of U:G mismatches by activation-induced cytidine deaminase but differ in their subsequent mutagenic consequences. Although SHM relies on the generation of nondeleterious point mutations, CSR depends on the production of DNA double-strand breaks (DSBs) and their adequate recombination through nonhomologous end joining (NHEJ). MLH1, an ATPase member of the mismatch repair (MMR) machinery, is emerging as a likely regulator of whether a U:G mismatch progresses toward mutation or DSB formation. We conducted experiments on cancer modeled ATPase-deficient MLH1G67R knockin mice to determine the function that the ATPase domain of MLH1 mediates in SHM and CSR. Mlh1GR/GR mice displayed a significant decrease in CSR, mainly attributed to a reduction in the generation of DSBs and diminished accumulation of 53BP1 at the immunoglobulin switch regions. However, SHM was normal in these mice, which distinguishes MLH1 from upstream members of the MMR pathway and suggests a very specific role of its ATPase-dependent functions during CSR. In addition, we show that the residual switching events still taking place in Mlh1GR/GR mice display unique features, suggesting a role for the ATPase activity of MLH1 beyond the activation of the endonuclease functions of its MMR partner PMS2. A preference for switch junctions with longer microhomologies in Mlh1GR/GR mice suggests that through its ATPase activity, MLH1 also has an impact in DNA end processing, favoring canonical NHEJ downstream of the DSB. Collectively, our study shows that the ATPase domain of MLH1 is important to transmit the CSR signaling cascade both upstream and downstream of the generation of DSBs.

Original languageEnglish (US)
Pages (from-to)671-678
Number of pages8
JournalJournal of Experimental Medicine
Volume209
Issue number4
DOIs
StatePublished - Apr 9 2012

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Double-Stranded DNA Breaks
Genetic Recombination
Adenosine Triphosphatases
DNA Mismatch Repair
Immunoglobulin Switch Region
Endonucleases
Point Mutation
B-Lymphocytes
Mutation
Antibodies
DNA
Neoplasms

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Medicine(all)

Cite this

The ATPase activity of MLH1 is required to orchestrate DNA double-strand breaks and end processing during class switch recombination. / Chahwan, Richard; van Oers, Johanna M M; Avdievich, Elena; Zhao, Chunfang; Edelmann, Winfried; Scharff, Matthew D.; Roa, Sergio Roa.

In: Journal of Experimental Medicine, Vol. 209, No. 4, 09.04.2012, p. 671-678.

Research output: Contribution to journalArticle

Chahwan, Richard ; van Oers, Johanna M M ; Avdievich, Elena ; Zhao, Chunfang ; Edelmann, Winfried ; Scharff, Matthew D. ; Roa, Sergio Roa. / The ATPase activity of MLH1 is required to orchestrate DNA double-strand breaks and end processing during class switch recombination. In: Journal of Experimental Medicine. 2012 ; Vol. 209, No. 4. pp. 671-678.
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