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

AutorRuiz, José F. ; Lucas, Daniel; García-Palomero, Esther; Sáez, Ana I.; González, Manuel A.; Piris, Miguel Ángel; Bernad, Antonio; Blanco, Luis
Palabras claveDNA polymerase µ
Fecha de publicación1-nov-2004
EditorOxford University Press
CitaciónNucleic Acids Research 2004 32(19):5861-5873
ResumenDNA polymerase µ (Pol µ) 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 µ 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 µ 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 µ to mutation of G and C residues during SHM. In vitro analysis of Pol µ 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 µ to induce transient template/primer misalignments
Versión del editorhttp://dx.doi.org/10.1093/nar/gkh929
ISSN0305-1048 (print)
1362-4962 (online)
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