Article ; Online: Division of labor of Y-family polymerases in translesion-DNA synthesis for distinct types of DNA damage.
PloS one
2021 Volume 16, Issue 6, Page(s) e0252587
Abstract: ... is restarted via bypass replication by translesion DNA-synthesis polymerases, including the Y-family ... play complementary roles in bypassing MMS-induced damage. Our findings indicate that the three Y-family ...
Abstract | Living organisms are continuously under threat from a vast array of DNA-damaging agents, which impact genome DNA. DNA replication machinery stalls at damaged template DNA. The stalled replication fork is restarted via bypass replication by translesion DNA-synthesis polymerases, including the Y-family polymerases Polη, Polι, and Polκ, which possess the ability to incorporate nucleotides opposite the damaged template. To investigate the division of labor among these polymerases in vivo, we generated POLη-/-, POLι-/-, POLκ-/-, double knockout (KO), and triple knockout (TKO) mutants in all combinations from human TK6 cells. TKO cells exhibited a hypersensitivity to ultraviolet (UV), cisplatin (CDDP), and methyl methanesulfonate (MMS), confirming the pivotal role played by these polymerases in bypass replication of damaged template DNA. POLη-/- cells, but not POLι-/- or POLκ-/- cells, showed a strong sensitivity to UV and CDDP, while TKO cells showed a slightly higher sensitivity to UV and CDDP than did POLη-/- cells. On the other hand, TKO cells, but not all single KO cells, exhibited a significantly higher sensitivity to MMS than did wild-type cells. Consistently, DNA-fiber assay revealed that Polη plays a crucial role in bypassing lesions caused by UV-mimetic agent 4-nitroquinoline-1-oxide and CDDP, while all three polymerases play complementary roles in bypassing MMS-induced damage. Our findings indicate that the three Y-family polymerases play distinctly different roles in bypass replication, according to the type of DNA damage generated on the template strand. |
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MeSH term(s) | Cell Line ; Cisplatin/pharmacology ; DNA/genetics ; DNA/metabolism ; DNA Damage/drug effects ; DNA Damage/radiation effects ; DNA Repair ; DNA Replication ; DNA-Directed DNA Polymerase/deficiency ; DNA-Directed DNA Polymerase/genetics ; DNA-Directed DNA Polymerase/metabolism ; Gene Knockout Techniques ; Humans ; Methyl Methanesulfonate/pharmacology ; Ultraviolet Rays |
Chemical Substances | DNA (9007-49-2) ; Methyl Methanesulfonate (AT5C31J09G) ; DNA polymerase iota (EC 2.7.7.-) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; POLK protein, human (EC 2.7.7.7) ; Rad30 protein (EC 2.7.7.7) ; Cisplatin (Q20Q21Q62J) |
Language | English |
Publishing date | 2021-06-01 |
Publishing country | United States |
Document type | Journal Article ; Research Support, Non-U.S. Gov't |
ISSN | 1932-6203 |
ISSN (online) | 1932-6203 |
DOI | 10.1371/journal.pone.0252587 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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