Cargando…

Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing

Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda–sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A pre...

Descripción completa

Detalles Bibliográficos
Autores principales: Kim, Jin S., Nanfara, Michael T., Chodavarapu, Sundari, Jin, Kyeong S., Babu, Vignesh M. P., Ghazy, Mohamed A., Chung, Scisung, Kaguni, Jon M., Sutton, Mark D., Cho, Yunje
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5397184/
https://www.ncbi.nlm.nih.gov/pubmed/28168278
http://dx.doi.org/10.1093/nar/gkx081
_version_ 1783230220854099968
author Kim, Jin S.
Nanfara, Michael T.
Chodavarapu, Sundari
Jin, Kyeong S.
Babu, Vignesh M. P.
Ghazy, Mohamed A.
Chung, Scisung
Kaguni, Jon M.
Sutton, Mark D.
Cho, Yunje
author_facet Kim, Jin S.
Nanfara, Michael T.
Chodavarapu, Sundari
Jin, Kyeong S.
Babu, Vignesh M. P.
Ghazy, Mohamed A.
Chung, Scisung
Kaguni, Jon M.
Sutton, Mark D.
Cho, Yunje
author_sort Kim, Jin S.
collection PubMed
description Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda–sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A prediction is that the activity of Hda is tightly controlled to ensure that replication initiation occurs only once per cell cycle. Here, we determined the crystal structure of the Hda–β clamp complex. This complex contains two pairs of Hda dimers sandwiched between two β clamp rings to form an octamer that is stabilized by three discrete interfaces. Two separate surfaces of Hda make contact with the β clamp, which is essential for Hda function in RIDA. The third interface between Hda monomers occludes the active site arginine finger, blocking its access to DnaA. Taken together, our structural and mutational analyses of the Hda–β clamp complex indicate that the interaction of the β clamp with Hda controls the ability of Hda to interact with DnaA. In the octameric Hda–β clamp complex, the inability of Hda to interact with DnaA is a novel mechanism that may regulate Hda function.
format Online
Article
Text
id pubmed-5397184
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-53971842017-04-24 Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing Kim, Jin S. Nanfara, Michael T. Chodavarapu, Sundari Jin, Kyeong S. Babu, Vignesh M. P. Ghazy, Mohamed A. Chung, Scisung Kaguni, Jon M. Sutton, Mark D. Cho, Yunje Nucleic Acids Res Genome Integrity, Repair and Replication Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda–sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A prediction is that the activity of Hda is tightly controlled to ensure that replication initiation occurs only once per cell cycle. Here, we determined the crystal structure of the Hda–β clamp complex. This complex contains two pairs of Hda dimers sandwiched between two β clamp rings to form an octamer that is stabilized by three discrete interfaces. Two separate surfaces of Hda make contact with the β clamp, which is essential for Hda function in RIDA. The third interface between Hda monomers occludes the active site arginine finger, blocking its access to DnaA. Taken together, our structural and mutational analyses of the Hda–β clamp complex indicate that the interaction of the β clamp with Hda controls the ability of Hda to interact with DnaA. In the octameric Hda–β clamp complex, the inability of Hda to interact with DnaA is a novel mechanism that may regulate Hda function. Oxford University Press 2017-04-20 2017-02-07 /pmc/articles/PMC5397184/ /pubmed/28168278 http://dx.doi.org/10.1093/nar/gkx081 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Genome Integrity, Repair and Replication
Kim, Jin S.
Nanfara, Michael T.
Chodavarapu, Sundari
Jin, Kyeong S.
Babu, Vignesh M. P.
Ghazy, Mohamed A.
Chung, Scisung
Kaguni, Jon M.
Sutton, Mark D.
Cho, Yunje
Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title_full Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title_fullStr Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title_full_unstemmed Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title_short Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing
title_sort dynamic assembly of hda and the sliding clamp in the regulation of replication licensing
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5397184/
https://www.ncbi.nlm.nih.gov/pubmed/28168278
http://dx.doi.org/10.1093/nar/gkx081
work_keys_str_mv AT kimjins dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT nanfaramichaelt dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT chodavarapusundari dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT jinkyeongs dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT babuvigneshmp dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT ghazymohameda dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT chungscisung dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT kagunijonm dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT suttonmarkd dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing
AT choyunje dynamicassemblyofhdaandtheslidingclampintheregulationofreplicationlicensing