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Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis

DNA polymerase β (pol β) plays a central role in the DNA base excision repair pathway and also serves as an important model polymerase. Dynamic characterization of pol β from methyl-TROSY (13)C-(1)H multiple quantum CPMG relaxation dispersion experiments of Ile and Met sidechains and previous backbo...

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Autores principales: DeRose, Eugene F, Kirby, Thomas W, Mueller, Geoffrey A, Beard, William A, Wilson, Samuel H, London, Robert E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101544/
https://www.ncbi.nlm.nih.gov/pubmed/29917149
http://dx.doi.org/10.1093/nar/gky503
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author DeRose, Eugene F
Kirby, Thomas W
Mueller, Geoffrey A
Beard, William A
Wilson, Samuel H
London, Robert E
author_facet DeRose, Eugene F
Kirby, Thomas W
Mueller, Geoffrey A
Beard, William A
Wilson, Samuel H
London, Robert E
author_sort DeRose, Eugene F
collection PubMed
description DNA polymerase β (pol β) plays a central role in the DNA base excision repair pathway and also serves as an important model polymerase. Dynamic characterization of pol β from methyl-TROSY (13)C-(1)H multiple quantum CPMG relaxation dispersion experiments of Ile and Met sidechains and previous backbone relaxation dispersion measurements, reveals transitions in μs-ms dynamics in response to highly variable substrates. Recognition of a 1-nt-gapped DNA substrate is accompanied by significant backbone and sidechain motion in the lyase domain and the DNA binding subdomain of the polymerase domain, that may help to facilitate binding of the apoenzyme to the segments of the DNA upstream and downstream from the gap. Backbone μs-ms motion largely disappears after formation of the pol β–DNA complex, giving rise to an increase in uncoupled μs-ms sidechain motion throughout the enzyme. Formation of an abortive ternary complex using a non-hydrolyzable dNTP results in sidechain motions that fit to a single exchange process localized to the catalytic subdomain, suggesting that this motion may play a role in catalysis.
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spelling pubmed-61015442018-08-27 Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis DeRose, Eugene F Kirby, Thomas W Mueller, Geoffrey A Beard, William A Wilson, Samuel H London, Robert E Nucleic Acids Res Nucleic Acid Enzymes DNA polymerase β (pol β) plays a central role in the DNA base excision repair pathway and also serves as an important model polymerase. Dynamic characterization of pol β from methyl-TROSY (13)C-(1)H multiple quantum CPMG relaxation dispersion experiments of Ile and Met sidechains and previous backbone relaxation dispersion measurements, reveals transitions in μs-ms dynamics in response to highly variable substrates. Recognition of a 1-nt-gapped DNA substrate is accompanied by significant backbone and sidechain motion in the lyase domain and the DNA binding subdomain of the polymerase domain, that may help to facilitate binding of the apoenzyme to the segments of the DNA upstream and downstream from the gap. Backbone μs-ms motion largely disappears after formation of the pol β–DNA complex, giving rise to an increase in uncoupled μs-ms sidechain motion throughout the enzyme. Formation of an abortive ternary complex using a non-hydrolyzable dNTP results in sidechain motions that fit to a single exchange process localized to the catalytic subdomain, suggesting that this motion may play a role in catalysis. Oxford University Press 2018-08-21 2018-06-18 /pmc/articles/PMC6101544/ /pubmed/29917149 http://dx.doi.org/10.1093/nar/gky503 Text en Published by Oxford University Press on behalf of Nucleic Acids Research 2018. This work is written by (a) US Government employee(s) and is in the public domain in the US.
spellingShingle Nucleic Acid Enzymes
DeRose, Eugene F
Kirby, Thomas W
Mueller, Geoffrey A
Beard, William A
Wilson, Samuel H
London, Robert E
Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title_full Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title_fullStr Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title_full_unstemmed Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title_short Transitions in DNA polymerase β μs-ms dynamics related to substrate binding and catalysis
title_sort transitions in dna polymerase β μs-ms dynamics related to substrate binding and catalysis
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101544/
https://www.ncbi.nlm.nih.gov/pubmed/29917149
http://dx.doi.org/10.1093/nar/gky503
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