The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*

Clamp loaders load ring-shaped sliding clamps onto DNA. Once loaded onto DNA, sliding clamps bind to DNA polymerases to increase the processivity of DNA synthesis. To load clamps onto DNA, an open clamp loader-clamp complex must form. An unresolved question is whether clamp loaders capture clamps th...

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Main Authors:	Paschall, Christopher O., Thompson, Jennifer A., Marzahn, Melissa R., Chiraniya, Ankita, Hayner, Jaclyn N., O'Donnell, Mike, Robbins, Arthur H., McKenna, Robert, Bloom, Linda B.
Format:	Online
Language:	English
Published:	American Society for Biochemistry and Molecular Biology 2011
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3234947/

id	pubmed-3234947
recordtype	oai_dc
spelling	pubmed-32349472011-12-12 The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp* Paschall, Christopher O. Thompson, Jennifer A. Marzahn, Melissa R. Chiraniya, Ankita Hayner, Jaclyn N. O'Donnell, Mike Robbins, Arthur H. McKenna, Robert Bloom, Linda B. DNA and Chromosomes Clamp loaders load ring-shaped sliding clamps onto DNA. Once loaded onto DNA, sliding clamps bind to DNA polymerases to increase the processivity of DNA synthesis. To load clamps onto DNA, an open clamp loader-clamp complex must form. An unresolved question is whether clamp loaders capture clamps that have transiently opened or whether clamp loaders bind closed clamps and actively open clamps. A simple fluorescence-based clamp opening assay was developed to address this question and to determine how ATP binding contributes to clamp opening. A direct comparison of real time binding and opening reactions revealed that the Escherichia coli γ complex binds β first and then opens the clamp. Mutation of conserved “arginine fingers” in the γ complex that interact with bound ATP decreased clamp opening activity showing that arginine fingers make an important contribution to the ATP-induced conformational changes that allow the clamp loader to pry open the clamp. American Society for Biochemistry and Molecular Biology 2011-12-09 2011-10-04 /pmc/articles/PMC3234947/ /pubmed/21971175 http://dx.doi.org/10.1074/jbc.M111.268169 Text en © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Paschall, Christopher O. Thompson, Jennifer A. Marzahn, Melissa R. Chiraniya, Ankita Hayner, Jaclyn N. O'Donnell, Mike Robbins, Arthur H. McKenna, Robert Bloom, Linda B.
spellingShingle	Paschall, Christopher O. Thompson, Jennifer A. Marzahn, Melissa R. Chiraniya, Ankita Hayner, Jaclyn N. O'Donnell, Mike Robbins, Arthur H. McKenna, Robert Bloom, Linda B. The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
author_facet	Paschall, Christopher O. Thompson, Jennifer A. Marzahn, Melissa R. Chiraniya, Ankita Hayner, Jaclyn N. O'Donnell, Mike Robbins, Arthur H. McKenna, Robert Bloom, Linda B.
author_sort	Paschall, Christopher O.
title	The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
title_short	The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
title_full	The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
title_fullStr	The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
title_full_unstemmed	The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*
title_sort	escherichia coli clamp loader can actively pry open the β-sliding clamp*
description	Clamp loaders load ring-shaped sliding clamps onto DNA. Once loaded onto DNA, sliding clamps bind to DNA polymerases to increase the processivity of DNA synthesis. To load clamps onto DNA, an open clamp loader-clamp complex must form. An unresolved question is whether clamp loaders capture clamps that have transiently opened or whether clamp loaders bind closed clamps and actively open clamps. A simple fluorescence-based clamp opening assay was developed to address this question and to determine how ATP binding contributes to clamp opening. A direct comparison of real time binding and opening reactions revealed that the Escherichia coli γ complex binds β first and then opens the clamp. Mutation of conserved “arginine fingers” in the γ complex that interact with bound ATP decreased clamp opening activity showing that arginine fingers make an important contribution to the ATP-induced conformational changes that allow the clamp loader to pry open the clamp.
publisher	American Society for Biochemistry and Molecular Biology
publishDate	2011
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3234947/
_version_	1611493014620864512

The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*

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