Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA

Silvia Hormeno; Oliver J Wilkinson; Clara Aicart-Ramos; Sahiti Kuppa; Edwin Antony; Mark S Dillingham; Fernando Moreno-Herrero

doi:10.1073/pnas.2112376119

Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA

Silvia Hormeno, Oliver J Wilkinson, Clara Aicart-Ramos, Sahiti Kuppa, Edwin Antony, Mark S Dillingham^*, Fernando Moreno-Herrero^*

^*Corresponding author for this work

School of Biochemistry

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Abstract

SignificanceSingle-stranded DNA (ssDNA) is a key intermediate in many cellular DNA transactions, including DNA replication, repair, and recombination. Nascent ssDNA is rapidly bound by the Replication Protein A (RPA) complex, forming a nucleoprotein filament that both stabilizes ssDNA and mediates downstream processing events. Paradoxically, however, the very high affinity of RPA for ssDNA may block the recruitment of further factors. In this work, we show that RPA-ssDNA nucleoprotein filaments are specifically targeted by the human HELB helicase. Recruitment of HELB by RPA-ssDNA activates HELB translocation activity, leading to processive removal of upstream RPA complexes. This RPA clearance activity may underpin the diverse roles of HELB in replication and recombination.

Original language	English
Article number	e2112376119
Number of pages	12
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	119
Issue number	15
DOIs	https://doi.org/10.1073/pnas.2112376119
Publication status	Published - 6 Apr 2022

Bibliographical note

Funding Information:
Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio?S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer?Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit.

Funding Information:
ACKNOWLEDGMENTS. Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio–S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer–Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit.

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Copyright © 2022 the Author(s).

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@article{16369aae083b44edb5141ac20cce2e04,

title = "Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA",

abstract = "SignificanceSingle-stranded DNA (ssDNA) is a key intermediate in many cellular DNA transactions, including DNA replication, repair, and recombination. Nascent ssDNA is rapidly bound by the Replication Protein A (RPA) complex, forming a nucleoprotein filament that both stabilizes ssDNA and mediates downstream processing events. Paradoxically, however, the very high affinity of RPA for ssDNA may block the recruitment of further factors. In this work, we show that RPA-ssDNA nucleoprotein filaments are specifically targeted by the human HELB helicase. Recruitment of HELB by RPA-ssDNA activates HELB translocation activity, leading to processive removal of upstream RPA complexes. This RPA clearance activity may underpin the diverse roles of HELB in replication and recombination.",

author = "Silvia Hormeno and Wilkinson, {Oliver J} and Clara Aicart-Ramos and Sahiti Kuppa and Edwin Antony and Dillingham, {Mark S} and Fernando Moreno-Herrero",

note = "Funding Information: Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio?S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer?Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit. Funding Information: ACKNOWLEDGMENTS. Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio–S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer–Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit. Publisher Copyright: Copyright {\textcopyright} 2022 the Author(s).",

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Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA. / Hormeno, Silvia; Wilkinson, Oliver J; Aicart-Ramos, Clara et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 119, No. 15, e2112376119, 06.04.2022.

Research output: Contribution to journal › Article (Academic Journal) › peer-review

TY - JOUR

T1 - Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA

AU - Hormeno, Silvia

AU - Wilkinson, Oliver J

AU - Aicart-Ramos, Clara

AU - Kuppa, Sahiti

AU - Antony, Edwin

AU - Dillingham, Mark S

AU - Moreno-Herrero, Fernando

N1 - Funding Information: Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio?S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer?Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit. Funding Information: ACKNOWLEDGMENTS. Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/ FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/ 501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio–S2018/ NMT-4443 (to F.M.-H.) and NanoBioCancer–Y2018/BIO-4747 (to F.M.-H.). Unlabelled yeast (S. cerevisiae) RPA was a gift from Luke Yates, and BirA was a gift from Charles Grummit. Publisher Copyright: Copyright © 2022 the Author(s).

PY - 2022/4/6

Y1 - 2022/4/6

N2 - SignificanceSingle-stranded DNA (ssDNA) is a key intermediate in many cellular DNA transactions, including DNA replication, repair, and recombination. Nascent ssDNA is rapidly bound by the Replication Protein A (RPA) complex, forming a nucleoprotein filament that both stabilizes ssDNA and mediates downstream processing events. Paradoxically, however, the very high affinity of RPA for ssDNA may block the recruitment of further factors. In this work, we show that RPA-ssDNA nucleoprotein filaments are specifically targeted by the human HELB helicase. Recruitment of HELB by RPA-ssDNA activates HELB translocation activity, leading to processive removal of upstream RPA complexes. This RPA clearance activity may underpin the diverse roles of HELB in replication and recombination.

AB - SignificanceSingle-stranded DNA (ssDNA) is a key intermediate in many cellular DNA transactions, including DNA replication, repair, and recombination. Nascent ssDNA is rapidly bound by the Replication Protein A (RPA) complex, forming a nucleoprotein filament that both stabilizes ssDNA and mediates downstream processing events. Paradoxically, however, the very high affinity of RPA for ssDNA may block the recruitment of further factors. In this work, we show that RPA-ssDNA nucleoprotein filaments are specifically targeted by the human HELB helicase. Recruitment of HELB by RPA-ssDNA activates HELB translocation activity, leading to processive removal of upstream RPA complexes. This RPA clearance activity may underpin the diverse roles of HELB in replication and recombination.

U2 - 10.1073/pnas.2112376119

DO - 10.1073/pnas.2112376119

M3 - Article (Academic Journal)

C2 - 35385349

SN - 0027-8424

VL - 119

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 15

M1 - e2112376119

ER -

Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA

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