RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

Ummi B Abdullah; Joanna F McGouran; Sanja Brolih; Denis Ptchelkine; Afaf H El‐Sagheer; Tom Brown; Peter J McHugh

doi:10.15252/embj.201796664

RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

Ummi B Abdullah, Joanna F McGouran, Sanja Brolih, Denis Ptchelkine, Afaf H El‐Sagheer, Tom Brown, Peter J McHugh

Research output: Contribution to journal › Article › peer-review

45 Scopus citations

Abstract

During replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

Original language	English (US)
Pages (from-to)	2047-2060
Number of pages	14
Journal	The EMBO Journal
Volume	36
Issue number	14
DOIs	https://doi.org/10.15252/embj.201796664
State	Published - Jun 12 2017
Externally published	Yes

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: We thank our laboratory member Sook Y. Lee and Opher Gileadi (Structural Genomics Consortium, Oxford) for the recombinant SNM1A; Fumiko Esashi (Dunn School of Pathology, University of Oxford) and Marc Wold (Iowa State University, USA) for the RPA and RPA70ΔC442, respectively; Hannah Baddock (WIMM), Hazel Aitkenhead and Joseph A. Newman (Structural Genomics Consortium, University of Oxford) for assistance with fluorescence anisotropy. P.J.M. was supported by MRC (Medical Research Council, grant MR/L007665/1). A.H.E.S. and T.B. were supported by BBSRC (Biotechnology and Biological Sciences Research Council, grant BB/J001694/1) BBSRC: sLOLA grant BB/J001694/1 “Extending the boundaries of nucleic acid chemistry” J.F.M. was supported by King Abdullah University of Science and Technology (KAUST) for a grant awarded to B. Nordén. U.B.A. was supported by the Malaysia's King Scholarship (Biasiswa Yang Di-Pertuan Agong).
This publication acknowledges KAUST support, but has no KAUST affiliated authors.

Access to Document

10.15252/embj.201796664

Cite this

@article{18719c179ff0439abf8f503fe88c8cee,

title = "RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks",

abstract = "During replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.",

author = "Abdullah, {Ummi B} and McGouran, {Joanna F} and Sanja Brolih and Denis Ptchelkine and El‐Sagheer, {Afaf H} and Tom Brown and McHugh, {Peter J}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank our laboratory member Sook Y. Lee and Opher Gileadi (Structural Genomics Consortium, Oxford) for the recombinant SNM1A; Fumiko Esashi (Dunn School of Pathology, University of Oxford) and Marc Wold (Iowa State University, USA) for the RPA and RPA70ΔC442, respectively; Hannah Baddock (WIMM), Hazel Aitkenhead and Joseph A. Newman (Structural Genomics Consortium, University of Oxford) for assistance with fluorescence anisotropy. P.J.M. was supported by MRC (Medical Research Council, grant MR/L007665/1). A.H.E.S. and T.B. were supported by BBSRC (Biotechnology and Biological Sciences Research Council, grant BB/J001694/1) BBSRC: sLOLA grant BB/J001694/1 “Extending the boundaries of nucleic acid chemistry” J.F.M. was supported by King Abdullah University of Science and Technology (KAUST) for a grant awarded to B. Nord{\'e}n. U.B.A. was supported by the Malaysia's King Scholarship (Biasiswa Yang Di-Pertuan Agong). This publication acknowledges KAUST support, but has no KAUST affiliated authors.",

year = "2017",

month = jun,

day = "12",

doi = "10.15252/embj.201796664",

language = "English (US)",

volume = "36",

pages = "2047--2060",

journal = "The EMBO Journal",

issn = "0261-4189",

publisher = "EMBO Press",

number = "14",

}

TY - JOUR

T1 - RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

AU - Abdullah, Ummi B

AU - McGouran, Joanna F

AU - Brolih, Sanja

AU - Ptchelkine, Denis

AU - El‐Sagheer, Afaf H

AU - Brown, Tom

AU - McHugh, Peter J

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank our laboratory member Sook Y. Lee and Opher Gileadi (Structural Genomics Consortium, Oxford) for the recombinant SNM1A; Fumiko Esashi (Dunn School of Pathology, University of Oxford) and Marc Wold (Iowa State University, USA) for the RPA and RPA70ΔC442, respectively; Hannah Baddock (WIMM), Hazel Aitkenhead and Joseph A. Newman (Structural Genomics Consortium, University of Oxford) for assistance with fluorescence anisotropy. P.J.M. was supported by MRC (Medical Research Council, grant MR/L007665/1). A.H.E.S. and T.B. were supported by BBSRC (Biotechnology and Biological Sciences Research Council, grant BB/J001694/1) BBSRC: sLOLA grant BB/J001694/1 “Extending the boundaries of nucleic acid chemistry” J.F.M. was supported by King Abdullah University of Science and Technology (KAUST) for a grant awarded to B. Nordén. U.B.A. was supported by the Malaysia's King Scholarship (Biasiswa Yang Di-Pertuan Agong). This publication acknowledges KAUST support, but has no KAUST affiliated authors.

PY - 2017/6/12

Y1 - 2017/6/12

N2 - During replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

AB - During replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

UR - http://hdl.handle.net/10754/625123

UR - https://onlinelibrary.wiley.com/doi/abs/10.15252/embj.201796664

UR - http://www.scopus.com/inward/record.url?scp=85020408671&partnerID=8YFLogxK

U2 - 10.15252/embj.201796664

DO - 10.15252/embj.201796664

M3 - Article

SN - 0261-4189

VL - 36

SP - 2047

EP - 2060

JO - The EMBO Journal

JF - The EMBO Journal

IS - 14

ER -

RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Cite this