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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Veröffentlicht in:Nucleic acids research 2017-11, Vol.45 (20), p.12025-12038
Hauptverfasser: Ahdash, Zainab, Lau, Andy M, Byrne, Robert Thomas, Lammens, Katja, Stüetzer, Alexandra, Urlaub, Henning, Booth, Paula J, Reading, Eamonn, Hopfner, Karl-Peter, Politis, Argyris
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container_end_page 12038
container_issue 20
container_start_page 12025
container_title Nucleic acids research
container_volume 45
creator Ahdash, Zainab
Lau, Andy M
Byrne, Robert Thomas
Lammens, Katja
Stüetzer, Alexandra
Urlaub, Henning
Booth, Paula J
Reading, Eamonn
Hopfner, Karl-Peter
Politis, Argyris
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.
doi_str_mv 10.1093/nar/gkx890
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subjects Adenosine Triphosphate - chemistry
Adenosine Triphosphate - metabolism
Archaeal Proteins - chemistry
Archaeal Proteins - genetics
Archaeal Proteins - metabolism
Binding Sites - genetics
Deoxyribonucleases - chemistry
Deoxyribonucleases - genetics
Deoxyribonucleases - metabolism
DNA Breaks, Double-Stranded
DNA Helicases - chemistry
DNA Helicases - genetics
DNA Helicases - metabolism
DNA Repair
DNA, Archaeal - chemistry
DNA, Archaeal - genetics
DNA, Archaeal - metabolism
Models, Molecular
Nucleic Acid Conformation
Protein Binding
Protein Domains
Protein Multimerization
Structural Biology
Sulfolobus solfataricus - enzymology
Sulfolobus solfataricus - genetics
Sulfolobus solfataricus - metabolism
title Mechanistic insight into the assembly of the HerA-NurA helicase-nuclease DNA end resection complex
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