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AID mutates E. coli suggesting a DNA deamination mechanism for antibody diversification

Nature volume 418, pages 99–104 (2002)Cite this article

Abstract

After gene rearrangement, immunoglobulin variable genes are diversified by somatic hypermutation or gene conversion, whereas the constant region is altered by class-switch recombination. All three processes depend on activation-induced cytidine deaminase (AID)1,2,3,4,5,6,7, a B-cell-specific protein that has been proposed (because of sequence homology1) to function by RNA editing. But indications that the three gene diversification processes might be initiated by a common type of DNA lesion8,9,10,11, together with the proposal that there is a first phase of hypermutation that targets dC/dG12, suggested to us that AID may function directly at dC/dG pairs. Here we show that expression of AID in Escherichia coli gives a mutator phenotype that yields nucleotide transitions at dC/dG in a context-dependent manner. Mutation triggered by AID is enhanced by a deficiency of uracil-DNA glycosylase, which indicates that AID functions by deaminating dC residues in DNA. We propose that diversification of functional immunoglobulin genes is triggered by AID-mediated deamination of dC residues in the immunoglobulin locus with the outcome—that is, hypermutation phases 1 and 2, gene conversion or switch recombination—dependent on the way in which the initiating dU/dG lesion is resolved.

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Figure 1: DNA deamination model of immunoglobulin gene diversification.
Figure 2: Expression of AID in E. coli yields a mutator phenotype that is enhanced by UDG deficiency.
Figure 3: Nature of the AID-induced mutations in an UDG-positive (ung+) background.
Figure 4: Nature of the AID-induced rpoB mutations in an UDG-deficient (ung-1) background.

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Acknowledgements

We thank A. Bhagwat, P. Holliger, R. Savva, B. Weiss and the Coli Genetic Stock Center for plasmids and bacterial strains; R. Beale and J. di Noia for discussions; J. Sale for comments on the manuscript; and the late César Milstein, a long-term colleague, for encouraging us to work on antibody diversification. S.K.P.-M. and R.S.H. were supported in part by EMBO and Burroughs Wellcome Fund Hitchings–Elion fellowships, respectively; this work was also supported in part by the Arthritis Research Campaign and Leukaemia Research Fund.

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Author notes

  1. Svend K. Petersen-Mahrt and Reuben S. Harris: These authors contributed equally to this work

  2. Michael S. Neuberger: Correspondence and requests for materials should be addressed to M.S.N.

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  1. Medical Research Council Laboratory of Molecular Biology, Cambridge, Hills Road, CB2 2QH, UK

    Svend K. Petersen-Mahrt, Reuben S. Harris & Michael S. Neuberger

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  1. Svend K. Petersen-Mahrt
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Correspondence to Michael S. Neuberger.

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Petersen-Mahrt, S., Harris, R. & Neuberger, M. AID mutates E. coli suggesting a DNA deamination mechanism for antibody diversification. Nature 418, 99–104 (2002). https://doi.org/10.1038/nature00862

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