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Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction

Clamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a...

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Autores principales: Subramanian, Subu, Gorday, Kent, Marcus, Kendra, Orellana, Matthew R, Ren, Peter, Luo, Xiao Ran, O'Donnell, Michael E, Kuriyan, John
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8121543/
https://www.ncbi.nlm.nih.gov/pubmed/33847559
http://dx.doi.org/10.7554/eLife.66181
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author Subramanian, Subu
Gorday, Kent
Marcus, Kendra
Orellana, Matthew R
Ren, Peter
Luo, Xiao Ran
O'Donnell, Michael E
Kuriyan, John
author_facet Subramanian, Subu
Gorday, Kent
Marcus, Kendra
Orellana, Matthew R
Ren, Peter
Luo, Xiao Ran
O'Donnell, Michael E
Kuriyan, John
author_sort Subramanian, Subu
collection PubMed
description Clamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a glutamine residue in the AAA+ module (Gln 118) that is not located at a catalytic or interfacial site. Gln 118 forms a hydrogen-bonded junction in a helical unit that we term the central coupler, because it connects the catalytic centers to DNA and the sliding clamp. A suppressor mutation indicates that hydrogen bonding in the junction is important, and molecular dynamics simulations reveal that it maintains rigidity in the central coupler. The glutamine-mediated junction is preserved in diverse AAA+ ATPases, suggesting that a connected network of hydrogen bonds that links ATP molecules is an essential aspect of allosteric communication in these proteins.
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spelling pubmed-81215432021-05-17 Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction Subramanian, Subu Gorday, Kent Marcus, Kendra Orellana, Matthew R Ren, Peter Luo, Xiao Ran O'Donnell, Michael E Kuriyan, John eLife Structural Biology and Molecular Biophysics Clamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a glutamine residue in the AAA+ module (Gln 118) that is not located at a catalytic or interfacial site. Gln 118 forms a hydrogen-bonded junction in a helical unit that we term the central coupler, because it connects the catalytic centers to DNA and the sliding clamp. A suppressor mutation indicates that hydrogen bonding in the junction is important, and molecular dynamics simulations reveal that it maintains rigidity in the central coupler. The glutamine-mediated junction is preserved in diverse AAA+ ATPases, suggesting that a connected network of hydrogen bonds that links ATP molecules is an essential aspect of allosteric communication in these proteins. eLife Sciences Publications, Ltd 2021-04-13 /pmc/articles/PMC8121543/ /pubmed/33847559 http://dx.doi.org/10.7554/eLife.66181 Text en © 2021, Subramanian et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Structural Biology and Molecular Biophysics
Subramanian, Subu
Gorday, Kent
Marcus, Kendra
Orellana, Matthew R
Ren, Peter
Luo, Xiao Ran
O'Donnell, Michael E
Kuriyan, John
Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title_full Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title_fullStr Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title_full_unstemmed Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title_short Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
title_sort allosteric communication in dna polymerase clamp loaders relies on a critical hydrogen-bonded junction
topic Structural Biology and Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8121543/
https://www.ncbi.nlm.nih.gov/pubmed/33847559
http://dx.doi.org/10.7554/eLife.66181
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