title: Mutational analysis of MukE reveals its role in focal subcellular localization of MukBEF.
abstract: Bacterial condensin MukBEF is essential for global folding of the Escherichia coli chromosome. MukB, a SMC (structural maintenance of chromosome) protein, comprises the core of this complex and is responsible for its ATP-modulated DNA binding and reshaping activities. MukF serves as a kleisin that modulates MukB-DNA interactions and links MukBs into macromolecular assemblies. Little is known about the function of MukE. Using random mutagenesis, we generated six loss-of-function point mutations in MukE. The surface mutations clustered in two places. One of them was at or close to the interface with MukF while the other was away from the known interactions of the protein. All loss-of-function mutations affected focal localization of MukBEF in live cells. In vitro, however, only some of them interfered with the assembly of MukBEF into a complex or the ability of MukEF to disrupt MukB-DNA interactions. Moreover, some MukE mutants were able to join intracellular foci formed by endogenous MukBEF and most of the mutants were efficiently incorporated into MukBEF even in the presence of endogenous MukE. These data reveal that focal localization of MukBEF involves other activities besides DNA binding and that MukE plays a central role in them.
authors.affiliation: Department of Chemistry and Biochemistry, University of Oklahoma, Norman, OK 73019, USA.
authors.forename: Weifeng
authors.initials: W
authors.lastname: She
authors.forename: Elena
authors.initials: E
authors.lastname: Mordukhova
authors.forename: Hang
authors.initials: H
authors.lastname: Zhao
authors.forename: Zoya M
authors.initials: ZM
authors.lastname: Petrushenko
authors.forename: Valentin V
authors.initials: VV
authors.lastname: Rybenkov
mesh_terms: D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000235:genetics; D050505:Mutant Proteins / Q000032:analysis / Q000235:genetics; D017354:Point Mutation; D011485:Protein Binding; D055503:Protein Multimerization; D012097:Repressor Proteins / Q000032:analysis* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D050505:Mutant Proteins; D012097:Repressor Proteins; C087115:mukE protein, E coli