Somatic hypermutation does not require Rad54 and Rad54B-mediated homologous recombination.
Secondary diversification of immunoglobulin (Ig) genes occurs through somatic hypermutation (SHM) in B cells of the germinal center (GC). The GC reaction is associated with a high frequency of DNA double-strand breaks (DSB) in the hypermutation domain of Ig genes. Homologous recombination (HR) is a prominent DSB repair pathway. Among the proteins involved in HR are the Rad-54 paralogues, Rad54 and Rad54B. To investigate whether Rad54/Rad54B-mediated HR is involved in SHM, we determined the ratio of mutated versus non-mutated Vlambda PCR products from memory (IgM-, IgD-, Vlambda1+) and GC (PNA(high), Vlambda1+) B cells, the mutation load, the mutation frequency, the base exchange pattern and the distribution of somatic mutations along the rearranged Vlambda light chain (VlambdaLC) genes. All these parameters of SHM were unaltered in memory and GC B cells lacking one or both Rad54 paralogues. Thus, our data indicate that Rad54 and Rad54B-mediated HR is not essential for SHM. In addition, the finding that the ablation of RAD51 paralogues causes an increase in SHM argues against a direct involvement of HR in promoting SHM.
Bross
Linda
L
Research Institute Growth and Development, Department of Immunology, University of Maastricht, Maastricht, The Netherlands.
Wesoly
Joanna
J
Buerstedde
Jean-Marie
JM
Kanaar
Roland
R
Jacobs
Heinz
H
D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry; D004265:DNA Helicases; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D005803:Genes, Immunoglobulin; D018858:Germinal Center / Q000737:chemistry; D007114:Immunization; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007146:Immunoglobulin lambda-Chains / Q000235:genetics; D007156:Immunologic Memory; D051379:Mice; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000172:deficiency / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000502:physiology*; D027041:Somatic Hypermutation, Immunoglobulin / Q000502:physiology*
D007135:Immunoglobulin Variable Region; D007146:Immunoglobulin lambda-Chains; D009687:Nuclear Proteins; C491221:RAD54B protein, human; D004265:DNA Helicases; C103534:Rad54l protein, mouse