Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing

Liu He; Rebecca Lever; Andrew Cubbon; Muhammad Tehseen; Tabitha Jenkins; Alice O Nottingham; Anya Horton; Hannah Betts; Martin Fisher; Samir Hamdan; Panos Soultanas; Edward L Bolt

doi:10.1093/nar/gkad032

Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing

Liu He, Rebecca Lever, Andrew Cubbon, Muhammad Tehseen, Tabitha Jenkins, Alice O Nottingham, Anya Horton, Hannah Betts, Martin Fisher, Samir Hamdan, Panos Soultanas, Edward L Bolt

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

5 Scopus citations

Abstract

DNA strand breaks are repaired by DNA synthesis from an exposed DNA end paired with a homologous DNA template. DNA polymerase delta (Pol δ) catalyses DNA synthesis in multiple eukaryotic DNA break repair pathways but triggers genome instability unless its activity is restrained. We show that human HelQ halts DNA synthesis by isolated Pol δ and Pol δ-PCNA-RPA holoenzyme. Using novel HelQ mutant proteins we identify that inhibition of Pol δ is independent of DNA binding, and maps to a 70 amino acid intrinsically disordered region of HelQ. Pol δ and its POLD3 subunit robustly stimulated DNA single-strand annealing by HelQ, and POLD3 and HelQ interact physically via the intrinsically disordered HelQ region. This data, and inability of HelQ to inhibit DNA synthesis by the POLD1 catalytic subunit of Pol δ, reveal a mechanism for limiting DNA synthesis and promoting DNA strand annealing during human DNA break repair, which centres on POLD3.

Original language	English (US)
Journal	Nucleic acids research
DOIs	https://doi.org/10.1093/nar/gkad032
State	Published - Jan 31 2023

Bibliographical note

KAUST Repository Item: Exported on 2023-02-08
Acknowledged KAUST grant number(s): BAS/1/1002-01-01
Acknowledgements: Nanna Therapeutics, Cambridge, U.K. (to E.L.B.); BBSRC [BB/T006625/1 to E.L.B., BB/R013357/1 to P.S.]; King Abdullah University of Science and Technology (KAUST) Baseline Funding [BAS/1/1002-01-01 to S.M.H.]; PhD studentships from the BBSRC DTP; University of Nottingham School of Chemistry. Funding for open access charge: University of Nottingham Gold. We thank Stuart Wood, Aurelio Reyes, Christopher Cooper and Stan Jozwiakowski for comments, and Michell Hawkins for gifting E. coli polymerase enzymes.

ASJC Scopus subject areas

Genetics

Access to Document

10.1093/nar/gkad032

https://repository.kaust.edu.sa/bitstream/10754/687551/1/gkad032.pdf

Cite this

@article{69dccab07bc64647bb5c990fa198453d,

title = "Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing",

abstract = "DNA strand breaks are repaired by DNA synthesis from an exposed DNA end paired with a homologous DNA template. DNA polymerase delta (Pol δ) catalyses DNA synthesis in multiple eukaryotic DNA break repair pathways but triggers genome instability unless its activity is restrained. We show that human HelQ halts DNA synthesis by isolated Pol δ and Pol δ-PCNA-RPA holoenzyme. Using novel HelQ mutant proteins we identify that inhibition of Pol δ is independent of DNA binding, and maps to a 70 amino acid intrinsically disordered region of HelQ. Pol δ and its POLD3 subunit robustly stimulated DNA single-strand annealing by HelQ, and POLD3 and HelQ interact physically via the intrinsically disordered HelQ region. This data, and inability of HelQ to inhibit DNA synthesis by the POLD1 catalytic subunit of Pol δ, reveal a mechanism for limiting DNA synthesis and promoting DNA strand annealing during human DNA break repair, which centres on POLD3.",

author = "Liu He and Rebecca Lever and Andrew Cubbon and Muhammad Tehseen and Tabitha Jenkins and Alice O Nottingham and Anya Horton and Hannah Betts and Martin Fisher and Samir Hamdan and Panos Soultanas and Edward L Bolt",

note = "KAUST Repository Item: Exported on 2023-02-08 Acknowledged KAUST grant number(s): BAS/1/1002-01-01 Acknowledgements: Nanna Therapeutics, Cambridge, U.K. (to E.L.B.); BBSRC [BB/T006625/1 to E.L.B., BB/R013357/1 to P.S.]; King Abdullah University of Science and Technology (KAUST) Baseline Funding [BAS/1/1002-01-01 to S.M.H.]; PhD studentships from the BBSRC DTP; University of Nottingham School of Chemistry. Funding for open access charge: University of Nottingham Gold. We thank Stuart Wood, Aurelio Reyes, Christopher Cooper and Stan Jozwiakowski for comments, and Michell Hawkins for gifting E. coli polymerase enzymes.",

year = "2023",

month = jan,

day = "31",

doi = "10.1093/nar/gkad032",

language = "English (US)",

journal = "Nucleic acids research",

issn = "0305-1048",

publisher = "OXFORD UNIV PRESS",

}

TY - JOUR

T1 - Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing

AU - He, Liu

AU - Lever, Rebecca

AU - Cubbon, Andrew

AU - Tehseen, Muhammad

AU - Jenkins, Tabitha

AU - Nottingham, Alice O

AU - Horton, Anya

AU - Betts, Hannah

AU - Fisher, Martin

AU - Hamdan, Samir

AU - Soultanas, Panos

AU - Bolt, Edward L

N1 - KAUST Repository Item: Exported on 2023-02-08 Acknowledged KAUST grant number(s): BAS/1/1002-01-01 Acknowledgements: Nanna Therapeutics, Cambridge, U.K. (to E.L.B.); BBSRC [BB/T006625/1 to E.L.B., BB/R013357/1 to P.S.]; King Abdullah University of Science and Technology (KAUST) Baseline Funding [BAS/1/1002-01-01 to S.M.H.]; PhD studentships from the BBSRC DTP; University of Nottingham School of Chemistry. Funding for open access charge: University of Nottingham Gold. We thank Stuart Wood, Aurelio Reyes, Christopher Cooper and Stan Jozwiakowski for comments, and Michell Hawkins for gifting E. coli polymerase enzymes.

PY - 2023/1/31

Y1 - 2023/1/31

N2 - DNA strand breaks are repaired by DNA synthesis from an exposed DNA end paired with a homologous DNA template. DNA polymerase delta (Pol δ) catalyses DNA synthesis in multiple eukaryotic DNA break repair pathways but triggers genome instability unless its activity is restrained. We show that human HelQ halts DNA synthesis by isolated Pol δ and Pol δ-PCNA-RPA holoenzyme. Using novel HelQ mutant proteins we identify that inhibition of Pol δ is independent of DNA binding, and maps to a 70 amino acid intrinsically disordered region of HelQ. Pol δ and its POLD3 subunit robustly stimulated DNA single-strand annealing by HelQ, and POLD3 and HelQ interact physically via the intrinsically disordered HelQ region. This data, and inability of HelQ to inhibit DNA synthesis by the POLD1 catalytic subunit of Pol δ, reveal a mechanism for limiting DNA synthesis and promoting DNA strand annealing during human DNA break repair, which centres on POLD3.

AB - DNA strand breaks are repaired by DNA synthesis from an exposed DNA end paired with a homologous DNA template. DNA polymerase delta (Pol δ) catalyses DNA synthesis in multiple eukaryotic DNA break repair pathways but triggers genome instability unless its activity is restrained. We show that human HelQ halts DNA synthesis by isolated Pol δ and Pol δ-PCNA-RPA holoenzyme. Using novel HelQ mutant proteins we identify that inhibition of Pol δ is independent of DNA binding, and maps to a 70 amino acid intrinsically disordered region of HelQ. Pol δ and its POLD3 subunit robustly stimulated DNA single-strand annealing by HelQ, and POLD3 and HelQ interact physically via the intrinsically disordered HelQ region. This data, and inability of HelQ to inhibit DNA synthesis by the POLD1 catalytic subunit of Pol δ, reveal a mechanism for limiting DNA synthesis and promoting DNA strand annealing during human DNA break repair, which centres on POLD3.

UR - http://hdl.handle.net/10754/687551

UR - https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkad032/7016441

U2 - 10.1093/nar/gkad032

DO - 10.1093/nar/gkad032

M3 - Article

C2 - 36718939

SN - 0305-1048

JO - Nucleic acids research

JF - Nucleic acids research

ER -

Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing

Abstract

Bibliographical note

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this