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

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

doi:10.1093/nar/gkad032

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

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

Authors

Liu He

Rebecca Lever

Andrew Cubbon

Muhammad Tehseen

Tabitha Jenkins

Alice O. Nottingham

Anya Horton

Hannah Betts

Martin Fisher

Samir M. Hamdan

PANOS SOULTANAS PANOS.SOULTANAS@NOTTINGHAM.AC.UK
Professor of Biological Chemistry

ED BOLT ED.BOLT@NOTTINGHAM.AC.UK
Professor of Molecular Biology

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.

Citation

He, L., Lever, R., Cubbon, A., Tehseen, M., Jenkins, T., Nottingham, A. O., …Bolt, E. L. (2023). Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing. Nucleic Acids Research, 51(4), 1740-1749. https://doi.org/10.1093/nar/gkad032

Journal Article Type	Article
Acceptance Date	Jan 10, 2023
Online Publication Date	Jan 31, 2023
Publication Date	Feb 28, 2023
Deposit Date	Feb 6, 2023
Publicly Available Date	Feb 7, 2023
Journal	Nucleic acids research
Print ISSN	0305-1048
Electronic ISSN	1362-4962
Publisher	Oxford University Press (OUP)
Peer Reviewed	Peer Reviewed
Volume	51
Issue	4
Pages	1740-1749
DOI	https://doi.org/10.1093/nar/gkad032
Keywords	Genetics
Public URL	https://nottingham-repository.worktribe.com/output/16802154
Publisher URL	https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkad032/7016441

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.