title: Further evidence for involvement of a noncanonical function of uracil DNA glycosylase in class switch recombination.
abstract: Activation-induced cytidine deaminase (AID) introduces DNA cleavage in the Ig gene locus to initiate somatic hypermutation (SHM) and class switch recombination (CSR) in B cells. The DNA deamination model assumes that AID deaminates cytidine (C) on DNA and generates uridine (U), resulting in DNA cleavage after removal of U by uracil DNA glycosylase (UNG). Although UNG deficiency reduces CSR efficiency to one tenth, we reported that catalytically inactive mutants of UNG were fully proficient in CSR and that several mutants at noncatalytic sites lost CSR activity, indicating that enzymatic activity of UNG is not required for CSR. In this report we show that CSR activity by many UNG mutants critically depends on its N-terminal domain, irrespective of their enzymatic activities. Dissociation of the catalytic and CSR activity was also found in another UNG family member, SMUG1, and its mutants. We also show that Ugi, a specific peptide inhibitor of UNG, inhibits CSR without reducing DNA cleavage of the S (switch) region, confirming dispensability of UNG in DNA cleavage in CSR. It is therefore likely that UNG is involved in a repair step after DNA cleavage in CSR. Furthermore, requirement of the N terminus but not enzymatic activity of UNG mutants for CSR indicates that the UNG protein structure is critical. The present findings support our earlier proposal that CSR depends on a noncanonical function of the UNG protein (e.g., as a scaffold for repair enzymes) that might be required for the recombination reaction after DNA cleavage.
authors.affiliation: Department of Immunology and Genomic Medicine, Graduate School of Medicine, Kyoto University, Yoshida Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Nasim A
authors.initials: NA
authors.lastname: Begum
authors.forename: Andre
authors.initials: A
authors.lastname: Stanlie
authors.forename: Tomomitsu
authors.initials: T
authors.lastname: Doi
authors.forename: Yoko
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Hai Wei
authors.initials: HW
authors.lastname: Jin
authors.forename: Yong Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nagaoka
authors.forename: Tasuku
authors.initials: T
authors.lastname: Honjo
mesh_terms: D000818:Animals; D001483:Base Sequence; D055162:Biocatalysis; D002460:Cell Line; D017931:DNA Primers; D017578:Immunoglobulin Class Switching; D051379:Mice; D009154:Mutation; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D051981:Uracil-DNA Glycosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D017931:DNA Primers; D051981:Uracil-DNA Glycosidase