Distinct architectural requirements for the <i>parS</i> centromeric sequence of the pSM19035 plasmid partition machinery.

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 <i>parS</i>-DNA and ParB form partition complexes that activate ParA-ATPase to drive the system dynamics. The essential <i>parS</i> 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 <i>parS_pSM</i> 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 <i>parS_pSM</i> 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.