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Ubc13 dosage is critical for immunoglobulin gene conversion and gene targeting in vertebrate cells.
Nucleic Acids Res. 38, 4701-4707 (2010)
In contrast to lower eukaryotes, most vertebrate cells are characterized by a moderate efficiency of homologous recombination (HR) and limited feasibility of targeted genetic modifications. As a notable exception, the chicken DT40 B cell line is distinguished by efficient homology-mediated repair of DNA lesions during Ig gene conversion, and also shows exceptionally high gene-targeting efficiencies. The molecular basis of these phenomena is elusive. Here we show that the activity levels of Ubc13, the E2 enzyme responsible for non-canonical K63-linked polyubiquitination, are critical for high efficiency of Ig gene conversion and gene targeting in DT40. Ubc13(+/-) cells show substantially lower homology-mediated repair, yet do not display changes in somatic hypermutation, overall DNA repair or cell proliferation. Our results suggest that modulation of the activity of K63-linked polyubiquitination may be used to customize HR efficiencies in vertebrate cells.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
CONJUGATING ENZYME UBC13; SOMATIC HYPERMUTATION; DNA-DAMAGE; UBIQUITIN; REPAIR; INTERMEDIATE; RECOGNITION; MECHANISMS; PROTEINS; BREAKS
ISSN (print) / ISBN
0305-1048
e-ISSN
1362-4962
Journal
Nucleic Acids Research
Quellenangaben
Volume: 38,
Issue: 14,
Pages: 4701-4707
Publisher
Oxford University Press
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Peer reviewed