The structure of the bacterial DNA segregation ATPase filament reveals the conformational plasticity of ParA upon DNA binding

TY - JOUR

T1 - The structure of the bacterial DNA segregation ATPase filament reveals the conformational plasticity of ParA upon DNA binding

AU - Parker, Alexandra V

AU - Mann, Daniel

AU - Tzokov, Svetomir B

AU - Hwang, Ling C

AU - Bergeron, Julien R C

N1 - Funding Information: A.V.P. was recipient of a PhD scholarship from the Global Strategic Alliance at the University of Sheffield. We are grateful to Dr Satpal Chodha for helpful discussion on ParA biochemistry D.M. was supported by BBSRC grant BB/R019061/1 (to J.R.C.B.). We acknowledge the University of Sheffield EM facility for assistance with negative-stain EM data collection, and cryo-EM grid screening. X-ray crystallography data for the ParA2vc apo and ADP-bound were collected at the Diamond Light Source (proposal MX24447), and the Cryo-EM data for the ParA2vc-DNA structure was collected at eBIC (proposal EM20970). Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/8/27

Y1 - 2021/8/27

N2 - The efficient segregation of replicated genetic material is an essential step for cell division. Bacterial cells use several evolutionarily-distinct genome segregation systems, the most common of which is the type I Par system. It consists of an adapter protein, ParB, that binds to the DNA cargo via interaction with the parS DNA sequence; and an ATPase, ParA, that binds nonspecific DNA and mediates cargo transport. However, the molecular details of how this system functions are not well understood. Here, we report the cryo-EM structure of the Vibrio cholerae ParA2 filament bound to DNA, as well as the crystal structures of this protein in various nucleotide states. These structures show that ParA forms a left-handed filament on DNA, stabilized by nucleotide binding, and that ParA undergoes profound structural rearrangements upon DNA binding and filament assembly. Collectively, our data suggest the structural basis for ParA's cooperative binding to DNA and the formation of high ParA density regions on the nucleoid.

AB - The efficient segregation of replicated genetic material is an essential step for cell division. Bacterial cells use several evolutionarily-distinct genome segregation systems, the most common of which is the type I Par system. It consists of an adapter protein, ParB, that binds to the DNA cargo via interaction with the parS DNA sequence; and an ATPase, ParA, that binds nonspecific DNA and mediates cargo transport. However, the molecular details of how this system functions are not well understood. Here, we report the cryo-EM structure of the Vibrio cholerae ParA2 filament bound to DNA, as well as the crystal structures of this protein in various nucleotide states. These structures show that ParA forms a left-handed filament on DNA, stabilized by nucleotide binding, and that ParA undergoes profound structural rearrangements upon DNA binding and filament assembly. Collectively, our data suggest the structural basis for ParA's cooperative binding to DNA and the formation of high ParA density regions on the nucleoid.

KW - Adenosine Triphosphatases/chemistry

KW - Bacterial Proteins/chemistry

KW - Chromosome Segregation

KW - Cryoelectron Microscopy

KW - Crystallography, X-Ray

KW - DNA, Bacterial/chemistry

KW - Nucleic Acid Conformation

KW - Protein Conformation

KW - Vibrio cholerae/chemistry

UR - http://www.scopus.com/inward/record.url?scp=85113776879&partnerID=8YFLogxK

U2 - 10.1038/s41467-021-25429-2

DO - 10.1038/s41467-021-25429-2

M3 - Article

C2 - 34453062

AN - SCOPUS:85113776879

SN - 2041-1723

VL - 12

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 5166

ER -