Cargando…

X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action

RuvB-Like transcription factors function in cell cycle regulation, development and human disease, such as cancer and heart hyperplasia. The mechanisms that regulate adenosine triphosphate (ATP)-dependent activity, oligomerization and post-translational modifications in this family of enzymes are yet...

Descripción completa

Detalles Bibliográficos
Autores principales: Silva, Sara T. N., Brito, José A., Arranz, Rocío, Sorzano, Carlos Óscar S., Ebel, Christine, Doutch, James, Tully, Mark D., Carazo, José-María, Carrascosa, José L., Matias, Pedro M., Bandeiras, Tiago M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6137109/
https://www.ncbi.nlm.nih.gov/pubmed/30213962
http://dx.doi.org/10.1038/s41598-018-31997-z
_version_ 1783355121166450688
author Silva, Sara T. N.
Brito, José A.
Arranz, Rocío
Sorzano, Carlos Óscar S.
Ebel, Christine
Doutch, James
Tully, Mark D.
Carazo, José-María
Carrascosa, José L.
Matias, Pedro M.
Bandeiras, Tiago M.
author_facet Silva, Sara T. N.
Brito, José A.
Arranz, Rocío
Sorzano, Carlos Óscar S.
Ebel, Christine
Doutch, James
Tully, Mark D.
Carazo, José-María
Carrascosa, José L.
Matias, Pedro M.
Bandeiras, Tiago M.
author_sort Silva, Sara T. N.
collection PubMed
description RuvB-Like transcription factors function in cell cycle regulation, development and human disease, such as cancer and heart hyperplasia. The mechanisms that regulate adenosine triphosphate (ATP)-dependent activity, oligomerization and post-translational modifications in this family of enzymes are yet unknown. We present the first crystallographic structure of full-length human RuvBL2 which provides novel insights into its mechanistic action and biology. The ring-shaped hexameric RuvBL2 structure presented here resolves for the first time the mobile domain II of the human protein, which is responsible for protein-protein interactions and ATPase activity regulation. Structural analysis suggests how ATP binding may lead to domain II motion through interactions with conserved N-terminal loop histidine residues. Furthermore, a comparison between hsRuvBL1 and 2 shows differences in surface charge distribution that may account for previously described differences in regulation. Analytical ultracentrifugation and cryo electron microscopy analyses performed on hsRuvBL2 highlight an oligomer plasticity that possibly reflects different physiological conformations of the protein in the cell, as well as that single-stranded DNA (ssDNA) can promote the oligomerization of monomeric hsRuvBL2. Based on these findings, we propose a mechanism for ATP binding and domain II conformational change coupling.
format Online
Article
Text
id pubmed-6137109
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-61371092018-09-15 X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action Silva, Sara T. N. Brito, José A. Arranz, Rocío Sorzano, Carlos Óscar S. Ebel, Christine Doutch, James Tully, Mark D. Carazo, José-María Carrascosa, José L. Matias, Pedro M. Bandeiras, Tiago M. Sci Rep Article RuvB-Like transcription factors function in cell cycle regulation, development and human disease, such as cancer and heart hyperplasia. The mechanisms that regulate adenosine triphosphate (ATP)-dependent activity, oligomerization and post-translational modifications in this family of enzymes are yet unknown. We present the first crystallographic structure of full-length human RuvBL2 which provides novel insights into its mechanistic action and biology. The ring-shaped hexameric RuvBL2 structure presented here resolves for the first time the mobile domain II of the human protein, which is responsible for protein-protein interactions and ATPase activity regulation. Structural analysis suggests how ATP binding may lead to domain II motion through interactions with conserved N-terminal loop histidine residues. Furthermore, a comparison between hsRuvBL1 and 2 shows differences in surface charge distribution that may account for previously described differences in regulation. Analytical ultracentrifugation and cryo electron microscopy analyses performed on hsRuvBL2 highlight an oligomer plasticity that possibly reflects different physiological conformations of the protein in the cell, as well as that single-stranded DNA (ssDNA) can promote the oligomerization of monomeric hsRuvBL2. Based on these findings, we propose a mechanism for ATP binding and domain II conformational change coupling. Nature Publishing Group UK 2018-09-13 /pmc/articles/PMC6137109/ /pubmed/30213962 http://dx.doi.org/10.1038/s41598-018-31997-z Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Silva, Sara T. N.
Brito, José A.
Arranz, Rocío
Sorzano, Carlos Óscar S.
Ebel, Christine
Doutch, James
Tully, Mark D.
Carazo, José-María
Carrascosa, José L.
Matias, Pedro M.
Bandeiras, Tiago M.
X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title_full X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title_fullStr X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title_full_unstemmed X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title_short X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action
title_sort x-ray structure of full-length human ruvb-like 2 – mechanistic insights into coupling between atp binding and mechanical action
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6137109/
https://www.ncbi.nlm.nih.gov/pubmed/30213962
http://dx.doi.org/10.1038/s41598-018-31997-z
work_keys_str_mv AT silvasaratn xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT britojosea xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT arranzrocio xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT sorzanocarlososcars xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT ebelchristine xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT doutchjames xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT tullymarkd xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT carazojosemaria xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT carrascosajosel xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT matiaspedrom xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction
AT bandeirastiagom xraystructureoffulllengthhumanruvblike2mechanisticinsightsintocouplingbetweenatpbindingandmechanicalaction