Cargando…

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

Human DNA helicase B (HELB) is a poorly characterized helicase suggested to play both positive and negative regulatory roles in DNA replication and recombination. In this work, we used bulk and single-molecule approaches to characterize the biochemical activities of HELB protein with a particular fo...

Descripción completa

Detalles Bibliográficos
Autores principales: Hormeno, Silvia, Wilkinson, Oliver J., Aicart-Ramos, Clara, Kuppa, Sahiti, Antony, Edwin, Dillingham, Mark S., Moreno-Herrero, Fernando
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9169624/
https://www.ncbi.nlm.nih.gov/pubmed/35385349
http://dx.doi.org/10.1073/pnas.2112376119
_version_ 1784721240285511680
author Hormeno, Silvia
Wilkinson, Oliver J.
Aicart-Ramos, Clara
Kuppa, Sahiti
Antony, Edwin
Dillingham, Mark S.
Moreno-Herrero, Fernando
author_facet Hormeno, Silvia
Wilkinson, Oliver J.
Aicart-Ramos, Clara
Kuppa, Sahiti
Antony, Edwin
Dillingham, Mark S.
Moreno-Herrero, Fernando
author_sort Hormeno, Silvia
collection PubMed
description Human DNA helicase B (HELB) is a poorly characterized helicase suggested to play both positive and negative regulatory roles in DNA replication and recombination. In this work, we used bulk and single-molecule approaches to characterize the biochemical activities of HELB protein with a particular focus on its interactions with Replication Protein A (RPA) and RPA–single-stranded DNA (ssDNA) filaments. HELB is a monomeric protein that binds tightly to ssDNA with a site size of ∼20 nucleotides. It couples ATP hydrolysis to translocation along ssDNA in the 5′ to 3′ direction accompanied by the formation of DNA loops. HELB also displays classical helicase activity, but this is very weak in the absence of an assisting force. HELB binds specifically to human RPA, which enhances its ATPase and ssDNA translocase activities but inhibits DNA unwinding. Direct observation of HELB on RPA nucleoprotein filaments shows that translocating HELB concomitantly clears RPA from ssDNA. This activity, which can allow other proteins access to ssDNA intermediates despite their shielding by RPA, may underpin the diverse roles of HELB in cellular DNA transactions.
format Online
Article
Text
id pubmed-9169624
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher National Academy of Sciences
record_format MEDLINE/PubMed
spelling pubmed-91696242022-06-07 Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA Hormeno, Silvia Wilkinson, Oliver J. Aicart-Ramos, Clara Kuppa, Sahiti Antony, Edwin Dillingham, Mark S. Moreno-Herrero, Fernando Proc Natl Acad Sci U S A Biological Sciences Human DNA helicase B (HELB) is a poorly characterized helicase suggested to play both positive and negative regulatory roles in DNA replication and recombination. In this work, we used bulk and single-molecule approaches to characterize the biochemical activities of HELB protein with a particular focus on its interactions with Replication Protein A (RPA) and RPA–single-stranded DNA (ssDNA) filaments. HELB is a monomeric protein that binds tightly to ssDNA with a site size of ∼20 nucleotides. It couples ATP hydrolysis to translocation along ssDNA in the 5′ to 3′ direction accompanied by the formation of DNA loops. HELB also displays classical helicase activity, but this is very weak in the absence of an assisting force. HELB binds specifically to human RPA, which enhances its ATPase and ssDNA translocase activities but inhibits DNA unwinding. Direct observation of HELB on RPA nucleoprotein filaments shows that translocating HELB concomitantly clears RPA from ssDNA. This activity, which can allow other proteins access to ssDNA intermediates despite their shielding by RPA, may underpin the diverse roles of HELB in cellular DNA transactions. National Academy of Sciences 2022-04-06 2022-04-12 /pmc/articles/PMC9169624/ /pubmed/35385349 http://dx.doi.org/10.1073/pnas.2112376119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Biological Sciences
Hormeno, Silvia
Wilkinson, Oliver J.
Aicart-Ramos, Clara
Kuppa, Sahiti
Antony, Edwin
Dillingham, Mark S.
Moreno-Herrero, Fernando
Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title_full Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title_fullStr Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title_full_unstemmed Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title_short Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA
title_sort human helb is a processive motor protein that catalyzes rpa clearance from single-stranded dna
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9169624/
https://www.ncbi.nlm.nih.gov/pubmed/35385349
http://dx.doi.org/10.1073/pnas.2112376119
work_keys_str_mv AT hormenosilvia humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT wilkinsonoliverj humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT aicartramosclara humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT kuppasahiti humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT antonyedwin humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT dillinghammarks humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna
AT morenoherrerofernando humanhelbisaprocessivemotorproteinthatcatalyzesrpaclearancefromsinglestrandeddna