Pathways and signatures of mutagenesis at targeted DNA nicks

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....

Full description

Saved in:
Bibliographic Details
Published in:PLoS genetics 2021-04, Vol.17 (4), p.e1009329
Main Authors: Zhang, Yinbo, Davis, Luther, Maizels, Nancy
Format: Article
Language:English
Subjects:
Asymmetry
Biology and life sciences
BRCA2 protein
BRCA2 Protein - genetics
Cell Cycle - genetics
Deoxyribonucleic acid
DNA
DNA Breaks, Double-Stranded
DNA Breaks, Single-Stranded
DNA damage
DNA Damage - genetics
DNA Helicases - genetics
DNA Polymerase theta
DNA repair
DNA Repair - genetics
DNA-Binding Proteins - genetics
DNA-Directed DNA Polymerase - genetics
Experiments
Gene expression
Genetic research
High-Throughput Nucleotide Sequencing
Humans
INDEL Mutation - genetics
Ionizing radiation
Mad2 Proteins - genetics
Mutagenesis
Mutagenesis - genetics
Mutation
Nuclease
Nucleotidyltransferases - genetics
Observations
Oligonucleotides
Research and Analysis Methods
RNA, Guide, Kinetoplastida - genetics
Signal Transduction - genetics
Transcription
Tumors
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1553-7404
1553-7390
1553-7404
DOI:10.1371/journal.pgen.1009329