Regulation of class switch recombination and somatic mutation by AID phosphorylation.
Activation-induced cytidine deaminase (AID) is a mutator enzyme that initiates somatic mutation and class switch recombination in B lymphocytes by introducing uracil:guanine mismatches into DNA. Repair pathways process these mismatches to produce point mutations in the Ig variable region or double-stranded DNA breaks in the switch region DNA. However, AID can also produce off-target DNA damage, including mutations in oncogenes. Therefore, stringent regulation of AID is required for maintaining genomic stability during maturation of the antibody response. It has been proposed that AID phosphorylation at serine 38 (S38) regulates its activity, but this has not been tested in vivo. Using a combination of mass spectrometry and immunochemical approaches, we found that in addition to S38, AID is also phosphorylated at position threonine 140 (T140). Mutation of either S38 or T140 to alanine does not impact catalytic activity, but interferes with class switching and somatic hypermutation in vivo. This effect is particularly pronounced in haploinsufficient mice where AID levels are limited. Although S38 is equally important for both processes, T140 phosphorylation preferentially affects somatic mutation, suggesting that posttranslational modification might contribute to the choice between hypermutation and class switching.
McBride
Kevin M
KM
Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10021, USA.
Gazumyan
Anna
A
Woo
Eileen M
EM
Schwickert
Tanja A
TA
Chait
Brian T
BT
Nussenzweig
Michel C
MC
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism / Q000502:physiology*; D003564:Cytidine Deaminase / Q000378:metabolism*; D017578:Immunoglobulin Class Switching / Q000502:physiology*; D007150:Immunohistochemistry; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D017354:Point Mutation / Q000235:genetics*; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase