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Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex

The HerA–NurA helicase–nuclease complex cooperates with Mre11 and Rad50 to coordinate the repair of double-stranded DNA breaks. Little is known, however, about the assembly mechanism and activation of the HerA–NurA. By combining hybrid mass spectrometry with cryo-EM, computational and biochemical da...

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Autores principales: Ahdash, Zainab, Lau, Andy M., Byrne, Robert Thomas, Lammens, Katja, Stüetzer, Alexandra, Urlaub, Henning, Booth, Paula J., Reading, Eamonn, Hopfner, Karl-Peter, Politis, Argyris
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5715905/
https://www.ncbi.nlm.nih.gov/pubmed/29149348
http://dx.doi.org/10.1093/nar/gkx890
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author Ahdash, Zainab
Lau, Andy M.
Byrne, Robert Thomas
Lammens, Katja
Stüetzer, Alexandra
Urlaub, Henning
Booth, Paula J.
Reading, Eamonn
Hopfner, Karl-Peter
Politis, Argyris
author_facet Ahdash, Zainab
Lau, Andy M.
Byrne, Robert Thomas
Lammens, Katja
Stüetzer, Alexandra
Urlaub, Henning
Booth, Paula J.
Reading, Eamonn
Hopfner, Karl-Peter
Politis, Argyris
author_sort Ahdash, Zainab
collection PubMed
description The HerA–NurA helicase–nuclease complex cooperates with Mre11 and Rad50 to coordinate the repair of double-stranded DNA breaks. Little is known, however, about the assembly mechanism and activation of the HerA–NurA. By combining hybrid mass spectrometry with cryo-EM, computational and biochemical data, we investigate the oligomeric formation of HerA and detail the mechanism of nucleotide binding to the HerA–NurA complex from thermophilic archaea. We reveal that ATP-free HerA and HerA-DNA complexes predominantly exist in solution as a heptamer and act as a DNA loading intermediate. The binding of either NurA or ATP stabilizes the hexameric HerA, indicating that HerA–NurA is activated by substrates and complex assembly. To examine the role of ATP in DNA translocation and processing, we investigated how nucleotides interact with the HerA–NurA. We show that while the hexameric HerA binds six nucleotides in an ‘all-or-none’ fashion, HerA–NurA harbors a highly coordinated pairwise binding mechanism and enables the translocation and processing of double-stranded DNA. Using molecular dynamics simulations, we reveal novel inter-residue interactions between the external ATP and the internal DNA binding sites. Overall, here we propose a stepwise assembly mechanism detailing the synergistic activation of HerA–NurA by ATP, which allows efficient processing of double-stranded DNA.
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spelling pubmed-57159052017-12-08 Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex Ahdash, Zainab Lau, Andy M. Byrne, Robert Thomas Lammens, Katja Stüetzer, Alexandra Urlaub, Henning Booth, Paula J. Reading, Eamonn Hopfner, Karl-Peter Politis, Argyris Nucleic Acids Res Structural Biology The HerA–NurA helicase–nuclease complex cooperates with Mre11 and Rad50 to coordinate the repair of double-stranded DNA breaks. Little is known, however, about the assembly mechanism and activation of the HerA–NurA. By combining hybrid mass spectrometry with cryo-EM, computational and biochemical data, we investigate the oligomeric formation of HerA and detail the mechanism of nucleotide binding to the HerA–NurA complex from thermophilic archaea. We reveal that ATP-free HerA and HerA-DNA complexes predominantly exist in solution as a heptamer and act as a DNA loading intermediate. The binding of either NurA or ATP stabilizes the hexameric HerA, indicating that HerA–NurA is activated by substrates and complex assembly. To examine the role of ATP in DNA translocation and processing, we investigated how nucleotides interact with the HerA–NurA. We show that while the hexameric HerA binds six nucleotides in an ‘all-or-none’ fashion, HerA–NurA harbors a highly coordinated pairwise binding mechanism and enables the translocation and processing of double-stranded DNA. Using molecular dynamics simulations, we reveal novel inter-residue interactions between the external ATP and the internal DNA binding sites. Overall, here we propose a stepwise assembly mechanism detailing the synergistic activation of HerA–NurA by ATP, which allows efficient processing of double-stranded DNA. Oxford University Press 2017-11-16 2017-10-09 /pmc/articles/PMC5715905/ /pubmed/29149348 http://dx.doi.org/10.1093/nar/gkx890 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ 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.
spellingShingle Structural Biology
Ahdash, Zainab
Lau, Andy M.
Byrne, Robert Thomas
Lammens, Katja
Stüetzer, Alexandra
Urlaub, Henning
Booth, Paula J.
Reading, Eamonn
Hopfner, Karl-Peter
Politis, Argyris
Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title_full Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title_fullStr Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title_full_unstemmed Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title_short Mechanistic insight into the assembly of the HerA–NurA helicase–nuclease DNA end resection complex
title_sort mechanistic insight into the assembly of the hera–nura helicase–nuclease dna end resection complex
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5715905/
https://www.ncbi.nlm.nih.gov/pubmed/29149348
http://dx.doi.org/10.1093/nar/gkx890
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