Distinct architectural requirements for the parS centromeric sequence of the pSM19035 plasmid partition machinery

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Abstract

Three-component ParABS partition systems ensure stable inheritance of many bacterial chromosomes and low-copy-number plasmids. ParA localizes to the nucleoid through its ATP-dependent non-specific DNA binding activity, whereas centromere-like parS-DNA and ParB form partition complexes that activate ParA-ATPase to drive the system dynamics. The essential parS sequence arrangements vary among ParABS systems, reflecting the architectural diversity of their partition complexes. Here, we focus on the pSM19035 plasmid partition system that uses a ParB_pSM of the ribbon-helix-helix (RHH) family. We show that parS_pSM with four or more contiguous ParB_pSM-binding sequence repeats is required to assemble a stable ParA_pSM-ParB_pSM complex and efficiently activate the ParA_pSM-ATPase, stimulating complex disassembly. Disruption of the contiguity of the parS_pSM sequence array destabilizes the ParA_pSM-ParB_pSM complex and prevents efficient ATPase activation. Our findings reveal the unique architecture of the pSM19035 partition complex and how it interacts with nucleoid-bound ParA_pSM-ATP.

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All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for all figures presented.

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Andrea Volante

National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, United States

Competing interests
The authors declare that no competing interests exist.
Juan Carlos Alonso

Department of Microbial Biotechnology, National Center for Biotechnology, Madrid, Spain

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5178-7179
Kiyoshi Mizuuchi

National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, United States

For correspondence
kiyoshimi@niddk.nih.gov

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-8193-9244

Funding

National Institute of Diabetes and Digestive and Kidney Diseases

Kiyoshi Mizuuchi

Ministerio de Ciencia e Innovación (2018-097054-B-I00,2021AEP031)

Juan Carlos Alonso

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.