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

DOI: 10.1038/s41598-018-31997-z DOI Help

Authors: Sara Silva (Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa) , Jose A. Brito (Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa) , Rocío Arranz (Centro Nacional de Biotecnología (CNB-CSIC)) , Carlos Óscar S. Sorzano (Centro Nacional de Biotecnología (CNB-CSIC)) , Christine Ebel (Univ. Grenoble Alpes, CNRS) , James Doutch (ISIS Pulsed Neutron and Muon Source) , Mark D. Tully (European Synchrotron Radiation Facility (ESRF)) , José-maría Carazo (Centro Nacional de Biotecnología (CNB-CSIC)) , José L. Carrascosa (Centro Nacional de Biotecnología (CNB-CSIC)) , Pedro M. Matias (Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa; iBET, Instituto de Biologia Experimental e Tecnológica) , Tiago M. Bandeiras (iBET, Instituto de Biologia Experimental e Tecnológica)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 8

State: Published (Approved)
Published: September 2018
Diamond Proposal Number(s): 13193

Open Access Open Access

Abstract: 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.

Subject Areas: Biology and Bio-materials


Instruments: B21-High Throughput SAXS

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s41598-018-31997-z.pdf