A scanning-to-incision switch in TFIIH-XPG induced by DNA damage licenses nucleotide excision repair

Bralić, Amer; Tehseen, Muhammad; Sobhy, Mohamed A.; Tsai, Chi-Lin; Alhudhali, Lubna; Yi, Gang; Yu, Jina; Yan, Chunli; Ivanov, Ivaylo; Tsutakawa, Susan E.; Tainer, John A; Hamdan, Samir M

doi:10.1093/nar/gkac1095

A scanning-to-incision switch in TFIIH-XPG induced by DNA damage licenses nucleotide excision repair

Journal Article · Thu Dec 08 04:00:00 EST 2022 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkac1095· OSTI ID:2283385

Bralić, Amer ^[1]; Tehseen, Muhammad ^[1]; Sobhy, Mohamed A. ^[1]; Tsai, Chi-Lin ^[2]; Alhudhali, Lubna ^[1]; Yi, Gang ^[1]; Yu, Jina ^[3]; Yan, Chunli ^[3]; ^[3]; Tsutakawa, Susan E. ^[4]; ^[5]; ^[1]

King Abdullah University of Science and Technology (KAUST), Thuwal (Saudi Arabia)
University of Texas MD Anderson Cancer Center, Houston, TX (United States)
Georgia State Univ., Atlanta, GA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); University of Texas MD Anderson Cancer Center, Houston, TX (United States)

Nucleotide excision repair (NER) is critical for removing bulky DNA base lesions and avoiding diseases. NER couples lesion recognition by XPC to strand separation by XPB and XPD ATPases, followed by lesion excision by XPF and XPG nucleases. Here, we describe key regulatory mechanisms and roles of XPG for and beyond its cleavage activity. Strikingly, by combing single-molecule imaging and bulk cleavage assays, we found that XPG binding to the 7-subunit TFIIH core (coreTFIIH) stimulates coreTFIIH-dependent double-strand (ds)DNA unwinding 10-fold, and XPG-dependent DNA cleavage by up to 700-fold. Simultaneous monitoring of rates for coreTFIIH single-stranded (ss)DNA translocation and dsDNA unwinding showed XPG acts by switching ssDNA translocation to dsDNA unwinding as a likely committed step. Pertinent to the NER pathway regulation, XPG incision activity is suppressed during coreTFIIH translocation on DNA but is licensed when coreTFIIH stalls at the lesion or when ATP hydrolysis is blocked. Moreover, ≥15 nucleotides of 5'-ssDNA is a prerequisite for efficient translocation and incision. Our results unveil a paired coordination mechanism in which key lesion scanning and DNA incision steps are sequentially coordinated, and damaged patch removal is only licensed after generation of ≥15 nucleotides of 5'-ssDNA, ensuring the correct ssDNA bubble size before cleavage.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2283385

Journal Information:: Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 3 Vol. 51; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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