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

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 nonspecific DNA-binding activity, whereas centromere-like parS -DNA and ParB form partition complexes that activate...

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Published in:	eLife Vol. 11
Main Authors:	Volante, Andrea, Alonso, Juan Carlos, Mizuuchi, Kiyoshi
Format:	Journal Article
Language:	English
Published:	Cambridge eLife Sciences Publications Ltd 05-09-2022 eLife Sciences Publications, Ltd
Subjects:	Adenosine triphosphatase Asymmetry Biochemistry and Chemical Biology Cell Biology Cell division chromosome segregation Chromosomes Gene expression Heredity ParABS system plasmid partition Plasmids Proteins
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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 nonspecific 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.
AbstractList	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 nonspecific 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 ParBpSM of the ribbon-helix-helix (RHH) family. We show that parSpSM with four or more contiguous ParBpSM-binding sequence repeats is required to assemble a stable ParApSM-ParBpSM complex and efficiently activate the ParApSM-ATPase, stimulating complex disassembly. Disruption of the contiguity of the parSpSM sequence array destabilizes the ParApSM-ParBpSM complex and prevents efficient ATPase activation. Our findings reveal the unique architecture of the pSM19035 partition complex and how it interacts with nucleoid-bound ParApSM-ATP. 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 nonspecific 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.
Author	Alonso, Juan Carlos Volante, Andrea Mizuuchi, Kiyoshi
Author_xml	– sequence: 1 givenname: Andrea surname: Volante fullname: Volante, Andrea – sequence: 2 givenname: Juan Carlos orcidid: 0000-0002-5178-7179 surname: Alonso fullname: Alonso, Juan Carlos – sequence: 3 givenname: Kiyoshi orcidid: 0000-0001-8193-9244 surname: Mizuuchi fullname: Mizuuchi, Kiyoshi
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SubjectTerms	Adenosine triphosphatase Asymmetry Biochemistry and Chemical Biology Cell Biology Cell division chromosome segregation Chromosomes Gene expression Heredity ParABS system plasmid partition Plasmids Proteins
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Title	Distinct architectural requirements for the parS centromeric sequence of the pSM19035 plasmid partition machinery
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