Acyl carrier protein promotes MukBEF action in Escherichia coli chromosome organization-segregation

Josh P. Prince,Carol V. Robinson,David J. Sherratt,Jarno Mäkelä,Gemma L. M. Fisher,Jani R. Bolla,Lidia K. Arciszewska,Marjorie Fournier

doi:10.1038/s41467-021-27107-9

Abstract

Structural Maintenance of Chromosomes (SMC) complexes act ubiquitously to compact DNA linearly, thereby facilitating chromosome organization-segregation. SMC proteins have a conserved architecture, with a dimerization hinge and an ATPase head domain separated by a long antiparallel intramolecular coiled-coil. Dimeric SMC proteins interact with essential accessory proteins, kleisins that bridge the two subunits of an SMC dimer, and HAWK/KITE proteins that interact with kleisins. The ATPase activity of the Escherichia coli SMC protein, MukB, which is essential for its in vivo function, requires its interaction with the dimeric kleisin, MukF that in turn interacts with the KITE protein, MukE. Here we demonstrate that, in addition, MukB interacts specifically with Acyl Carrier Protein (AcpP) that has essential functions in fatty acid synthesis. We characterize the AcpP interaction at the joint of the MukB coiled-coil and show that the interaction is necessary for MukB ATPase and for MukBEF function in vivo.

Highlights

Structural Maintenance of Chromosomes (SMC) complexes act ubiquitously to compact DNA linearly, thereby facilitating chromosome organization-segregation
We propose that AcpP is an essential partner for MukBEF action in chromosome organization segregation
Analysis of samples containing AcpP and MukBN or MukBHN using native Mass Spectrometry revealed AcpP interacts with MukB with a 1:1 monomer-monomer stoichiometry (Fig. 1e, f), supporting data previously reported for Wild type (WT) MukB16

Summary

Introduction

Structural Maintenance of Chromosomes (SMC) complexes act ubiquitously to compact DNA linearly, thereby facilitating chromosome organization-segregation. The ATPase activity of the Escherichia coli SMC protein, MukB, which is essential for its in vivo function, requires its interaction with the dimeric kleisin, MukF that in turn interacts with the KITE protein, MukE. As with other SMC proteins, MukB dimers interact with their kleisin, MukF, through two distinct interaction sites; one in the “neck” region of the coiled-coils, located between the head and the joint of one monomer, and the “cap” region of the partner ATPase head (Fig. 1a)[5,14,15]. MukF dimerizes through an additional N-terminal dimeric winged-helix domain (WHD) This enables the joining of two dimeric MukBEF complexes into dimer of dimer (DoD) complexes that are essential for in vivo MukBEF function[6,15,16]. Because the lack of functional MukB can result in reduced proliferation in organisms that cause disease phenotypes, such as Vibrio vulnificus[21], MukB is a potential target for new antimicrobial therapeutics[22]

Methods

Results

Conclusion