Overexpression of human DNA polymerase μ (Pol μ) in a Burkitt's lymphoma cell line affects the somatic hypermutation rate

被引:33
作者
Ruiz, JF
Lucas, D
García-Palomero, E
Saez, AI
González, MA
Piris, MA
Bernad, A
Blanco, L
机构
[1] Univ Autonoma Madrid, CSIC, Ctr Biol Mol Severo Ochoa, Madrid 28049, Spain
[2] Univ Autonoma Madrid, CSIC, Ctr Nacl Biotecnol, Dept Inmunol & Oncol, Madrid 28049, Spain
[3] Ctr Nacl Invest Oncol, Programa Patol Mol, Madrid, Spain
关键词
D O I
10.1093/nar/gkh929
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
DNA polymerase mu (Pol mu) is a DNA-dependent DNA polymerase closely related to terminal deoxynucleotidyl transferase (TdT), and prone to induce template/primer misalignments and misincorporation. In addition to a proposed general role in non-homologous end joining of double-strand breaks, its mutagenic potential and preferential expression in secondary lymphoid tissues support a role in somatic hypermutation (SHM) of immunoglobulin genes. Here, we show that human Pol mu protein is expressed in the nucleus of centroblasts obtained from human tonsils, forming a characteristic foci pattern resembling that of other DNA repair proteins in response to DNA damage. Overexpression of human Pol mu in Ramos cells, in which the SHM process is constitutive, augmented the somatic mutations specifically at the variable (V) region of the immunoglobulin genes. The nature of the mutations introduced, mostly base substitutions, supports the contribution of Pol mu to mutation of G and C residues during SHM. In vitro analysis of Pol mu misincorporation on specific templates, that mimic DNA repair intermediates and correspond to mutational hotspots, indicated that many of the mutations observed in vivo can be explained by the capacity of Pol mu to induce transient template/primer misalignments.
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收藏
页码:5861 / 5873
页数:13
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