Identification of murine B cell lines that undergo somatic hypermutation focused to A:T and G:C residues

Activation-induced deaminase (AID) is the master regulator of class switch recombination (CSR) and somatic hypermutation (SHM), but the mechanisms regulating AID function are obscure. The differential pattern of switch plasmid activity in three IgM+/AID+ and two IgG+/AID+ B cell lines prompted an analysis of global gene expression to discover the origin of these cells. Gene profiling suggested that the IgG+/AID+ B cell lines derived from germinal center B cells. Analysis of SHM potential demonstrates that the IgVκ domains are inducibly diversified at high rate during in vitro culture. The mutation spectra focused to A:T base pairs, revealing a component of the hypermutation program that occurs preferentially during phase 2 of SHM. The A:T error spectra were analyzed and were not characteristic of polymerase η activity. A differential pattern of three consensus motifs used for A:T base substitutions was observed in WT and Polη-, Msh2- and Msh6-deficient B cells. Strikingly, mutations in our B cell lines recapitulated the mutable motif profile for Polη and Msh2 deficiency, respectively, and suggest that an additional pathway for the generation of A:T mutations in SHM is conserved in mouse and human.