Pathways and signatures of mutagenesis at targeted DNA nicks

English version

Autoři: Yinbo Zhang ^aff001; Luther Davis ^aff001; Nancy Maizels ^aff001
Působiště autorů: Department of Immunology, University of Washington Medical School, Seattle, Washington, United States of America ^aff001; Department of Biochemistry, University of Washington Medical School, Seattle, Washington, United States of America ^aff002
Vyšlo v časopise: Pathways and signatures of mutagenesis at targeted DNA nicks. PLoS Genet 17(4): e1009329. doi:10.1371/journal.pgen.1009329
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1009329

Souhrn

Nicks are the most frequent form of DNA damage and a potential source of mutagenesis in human cells. By deep sequencing, we have identified factors and pathways that promote and limit mutagenic repair at a targeted nick in human cells. Mutations were distributed asymmetrically around the nick site. BRCA2 inhibited all categories of mutational events, including indels, SNVs and HDR. DNA2 and RPA promoted resection. DNA2 inhibited 1 bp deletions but contributed to longer deletions, as did REV7. POLQ stimulated SNVs. Parallel analysis of DSBs targeted to the same site identified similar roles for DNA2 and POLQ (but not REV7) in promoting deletions and for POLQ in stimulating SNVs. Insertions were infrequent at nicks, and most were 1 bp in length, as at DSBs. The translesion polymerase REV1 stimulated +1 insertions at one nick site but not another, illustrating the potential importance of sequence context in determining the outcome of mutagenic repair. These results highlight the potential for nicks to promote mutagenesis, especially in BRCA-deficient cells, and identify mutagenic signatures of DNA2, REV1, REV3, REV7 and POLQ.

Klíčová slova:

Cell cycle and cell division – DNA repair – G1 phase – Guide RNA – Insertion mutation – Mutagenesis – Recombinase polymerase amplification – Transfection

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