The AID antibody diversification enzyme is regulated by protein kinase A phosphorylation

• Published in: Nature Vol. 438; no. 7067; pp. 508 - 511
• Main Authors: Alt, Frederick W, Basu, Uttiya, Chaudhuri, Jayanta, Alpert, Craig, Dutt, Shilpee, Ranganath, Sheila, Li, Gang, Schrum, Jason Patrick, Manis, John P
• Format: Journal Article
• Language: English
• Published: London Nature Publishing 24.11.2005; Nature Publishing Group
• Subjects: Amino Acid Sequence; Animals; Antibodies, immunoglobulins; Antigens; B-Lymphocytes - enzymology; B-Lymphocytes - metabolism; Biological and medical sciences; Cell Line; Cells; Cyclic AMP-Dependent Protein Kinases - metabolism; Cytidine Deaminase - chemistry; Cytidine Deaminase - genetics; Cytidine Deaminase - metabolism; Deoxyribonucleic acid; DNA; Enzymes; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genetic recombination; Humans; Mice; Molecular immunology; Molecular Sequence Data; Mutation; Mutation - genetics; Phosphorylation; Protein Binding; Replication Protein A - metabolism; Somatic Hypermutation, Immunoglobulin; Phosphorylation; Enzyme; Protein kinase; Transferases
• ISSN: 0028-0836; 1476-4687; 1476-4679
• DOI: 10.1038/nature04255

Antibodies, which are produced by B-lineage cells, consist of immunoglobulin heavy (IgH) and light (IgL) chains that have amino-terminal variable regions and carboxy-terminal constant regions. In response to antigens, B cells undergo two types of genomic alterations to increase antibody diversity. Affinity for antigen can be increased by introduction of point mutations into IgH and IgL variable regions by somatic hypermutation. In addition, antibody effector functions can be altered by changing the expressed IgH constant region exons through IgH class switch recombination (CSR). Somatic hypermutation and CSR both require the B-cell-specific activation-induced cytidine deaminase protein (AID), which initiates these reactions through its single-stranded (ss)DNA-specific cytidine deaminase activity. In biochemical assays, replication protein A (RPA), a ssDNA-binding protein, associates with phosphorylated AID from activated B cells and enhances AID activity on transcribed double-stranded (ds)DNA containing somatic hypermutation or CSR target sequences. This AID-RPA association, which requires phosphorylation, may provide a mechanism for allowing AID to access dsDNA targets in activated B cells. Here we show that AID from B cells is phosphorylated on a consensus protein kinase A (PKA) site and that PKA is the physiological AID kinase. Thus, AID from non-lymphoid cells can be functionally phosphorylated by recombinant PKA to allow interaction with RPA and promote deamination of transcribed dsDNA substrates. Moreover, mutation of the major PKA phosphorylation site of AID preserves ssDNA deamination activity, but markedly reduces RPA-dependent dsDNA deamination activity and severely impairs the ability of AID to effect CSR in vivo. We conclude that PKA has a critical role in post-translational regulation of AID activity in B cells.