AID is required to initiate Nbs1/gamma-H2AX focus formation and mutations at sites of class switching
Authors: Thomas Ried
Leif Hanitsch
Bernardo Reina-San-Martin
Michael J Difilippantonio
Hua Tang Chen
William M Bonner
Simone Petersen
André Nussenzweig
Masamichi Muramatsuk
Tasuku Honjo
Christophe Redon
Patrick C Wilson
Michel C Nussenzweig
Rafael Casellas
Arkady Celeste
Duane R Pilch
Journal: Nature (2001)
Discipline: Multidisciplinary
Keywords: Class switch recombination; immunoglobulin; DNA recombination; heavy-chain constant region; variable region gene
Affiliations: Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA., Laboratory of Molecular Immunology, The Rockefeller University, and Howard Hughes Medical Institute, New York, New York 10021, USA., Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA., Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan., Laboratory of Molecular Pharmacology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

Abstract:  Class switch recombination (CSR) is a region-specific DNA recombination reaction that replaces one immunoglobulin heavy-chain constant region (Ch) gene with another. This enables a single variable (V) region gene to be used in conjunction with different downstream Ch genes, each having a unique biological activity. The molecular mechanisms that mediate CSR have not been defined, but activation-induced cytidine deaminase (AID), a putative RNA-editing enzyme, is required for this reaction. Here we report that the Nijmegen breakage syndrome protein (Nbs1) and phosphorylated H2A histone family member X (gamma-H2AX, also known as gamma-H2afx), which facilitate DNA double-strand break (DSB) repair, form nuclear foci at the Ch region in the G1 phase of the cell cycle in cells undergoing CSR, and that switching is impaired in H2AX-/- mice. Localization of Nbs1 and gamma-H2AX to the Igh locus during CSR is dependent on AID. In addition, AID is required for induction of switch region (S mu)-specific DNA lesions that precede CSR. These results place AID function upstream of the DNA modifications that initiate CSR. 

Retracted: No

PubPeer comments:
 Figure E9 appears to include a repeated image. *** ![file] , 
 #1 We apologize for this oversight. We have located previous raw data and revise the figure panel as attached here. Thanks for pointing it out ![file] , 
 #2 Thank you for responding and providing the corrected image here. Out of curiosity, will you also be contacting the journal? , 
 #3 #3 Yes, we are in the process of contacting the journal to submit the correction. Many thanks for pointing this out! , 
 Figure 1A. * Pink boxes: Two panels in the first column appear to shown an area of overlap. Since the rows are not labeled, it is not clear if these panels are supposed to represent samples from the same patients (n=3 while four rows are shown). The panels also do not correspond to the other three columns. Because the zoom factors are so different between the two panels, the scale bar on at least one of them appears incorrect. Could the authors please check? ![file] , 
 Thanks for pointing this out. The co-authors who provided the Figure 1A data confirmed that: 1) the IgG control experiment was performed in one sample instead of three samples since this is the technical negative control, and all of these antibodies are either Goat or Rabbit. So one IgG control is sufficient in this case; 2) the bottom row should have a scale bar of 25 um instead of 40um. We apologize for this oversight and will notify the journal accordingly. , 
 #6 Thank you. What is the status on the correction mentioned in #4? , 
 I contacted the journal a few times over last several months. I was told the journal is working on that. Thanks for your reminder. , 
 #8 Good news, the journal editor has responded: Hello. Thank you for alerting us to the PubPeer postings. We are currently working on a response, and hope to either post or publish that response shortly. Please let me know if you need any further information at this time. , 
 Yes. Thank you. For the Figure 1A, the scale bar in the bottom row should be 20um after we reviewed earlier experimental data provided by our collaborator. Will revise it in the correction. Thanks all of your hard work to help us improve science. , 
 15 Dec 2020 erratum. https: www.atsjournals.org doi 10.1164 rccm.v202erratum8 The Journal has been alerted to errors in two figures in the article by Jiang and colleagues (1), published in the July 15, 2016, issue. In Figure 1A, an incorrect image was used for the lower left panel (Rb IgG, Goat IgG). In addition, the panels in the bottom row included scale bars that were incorrectly labeled; these should be 20 u03bcm instead of 40 u03bcm. A corrected version of the figure is included here. ,