TY - JOUR
T1 - Nucleotide and partner-protein control of bacterial replicative helicase structure and function
AU - Strycharska, Melania S.
AU - Arias-Palomo, Ernesto
AU - Lyubimov, Artem Y.
AU - Erzberger, Jan P.
AU - O'Shea, Valerie L.
AU - Bustamante, Carlos J.
AU - Berger, James M.
N1 - Funding Information:
The authors are grateful to the Nogales laboratory and the staff at beamlines 8.3.1 and 12.3.1, of the Advanced Light Source, and beamline BL4-2, of the Stanford Synchrotron Radiation Lightsource, for help with data collection and analysis, and to the Berger laboratory for helpful discussions and aid in preparing the manuscript. We also thank Richard Rymer for providing DnaG protein, Seychelle Vos for help collecting SAXS data, Gregory Hura for help with processing SAXS data, and Tiago Barros of the Kuriyan lab for the τ domains III and IV construct. This work was supported by the Programa Nacional de Movilidad de Recursos Humanos del Plan Nacional de Investigación from the Spanish Ministry of Education to E.A.-P., an American Cancer Society postdoctoral fellowship to A.Y.L., an NIH NRSA postdoctoral fellowship to V.L.O. (F32GM090803), and the NIGMS (to J.M.B., R01-GM071747).
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 2013/12/26
Y1 - 2013/12/26
N2 - Cellular replication forks are powered by ring-shaped, hexameric helicases that encircle and unwind DNA. To better understand the molecular mechanisms and control of these enzymes, we used multiple methods to investigate the bacterial replicative helicase, DnaB. A 3.3Å crystal structure of Aquifex aeolicus DnaB, complexed with nucleotide, reveals a newly discovered conformational state for this motor protein. Electron microscopy and small angle X-ray scattering studies confirm the state seen crystallographically, showing that the DnaB ATPase domains and an associated N-terminal collar transition between two physical states in a nucleotide-dependent manner. Mutant helicases locked in either collar state are active but display different capacities to support critical activities such as duplex translocation and primase-dependent RNA synthesis. Our findings establish the DnaB collar as an autoregulatory hub that controls the ability of the helicase to transition between different functional states in response to both nucleotide and replication initiation/elongation factors.
AB - Cellular replication forks are powered by ring-shaped, hexameric helicases that encircle and unwind DNA. To better understand the molecular mechanisms and control of these enzymes, we used multiple methods to investigate the bacterial replicative helicase, DnaB. A 3.3Å crystal structure of Aquifex aeolicus DnaB, complexed with nucleotide, reveals a newly discovered conformational state for this motor protein. Electron microscopy and small angle X-ray scattering studies confirm the state seen crystallographically, showing that the DnaB ATPase domains and an associated N-terminal collar transition between two physical states in a nucleotide-dependent manner. Mutant helicases locked in either collar state are active but display different capacities to support critical activities such as duplex translocation and primase-dependent RNA synthesis. Our findings establish the DnaB collar as an autoregulatory hub that controls the ability of the helicase to transition between different functional states in response to both nucleotide and replication initiation/elongation factors.
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U2 - 10.1016/j.molcel.2013.11.016
DO - 10.1016/j.molcel.2013.11.016
M3 - Article
C2 - 24373746
AN - SCOPUS:84891136745
VL - 52
SP - 844
EP - 854
JO - Molecular Cell
JF - Molecular Cell
SN - 1097-2765
IS - 6
ER -