title: Structural asymmetry in a conserved signaling system that regulates division, replication, and virulence of an intracellular pathogen.
abstract: We have functionally and structurally defined an essential protein phosphorelay that regulates expression of genes required for growth, division, and intracellular survival of the global zoonotic pathogen Brucella abortus. Our study delineates phosphoryl transfer through this molecular pathway, which initiates from the sensor kinase CckA and proceeds through the ChpT phosphotransferase to two regulatory substrates: CtrA and CpdR. Genetic perturbation of this system results in defects in cell growth and division site selection, and a specific viability deficit inside human phagocytic cells. Thus, proper control of B. abortus division site polarity is necessary for survival in the intracellular niche. We further define the structural foundations of signaling from the central phosphotransferase, ChpT, to its response regulator substrate, CtrA, and provide evidence that there are at least two modes of interaction between ChpT and CtrA, only one of which is competent to catalyze phosphoryltransfer. The structure and dynamics of the active site on each side of the ChpT homodimer are distinct, supporting a model in which quaternary structure of the 2:2 ChpT-CtrA complex enforces an asymmetric mechanism of phosphoryl transfer between ChpT and CtrA. Our study provides mechanistic understanding, from the cellular to the atomic scale, of a conserved transcriptional regulatory system that controls the cellular and infection biology of B. abortus. More generally, our results provide insight into the structural basis of two-component signal transduction, which is broadly conserved in bacteria, plants, and fungi.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; Howard Taylor Ricketts Laboratory, University of Chicago, Argonne National Laboratory, Argonne, IL 60439;
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Willett
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; Howard Taylor Ricketts Laboratory, University of Chicago, Argonne National Laboratory, Argonne, IL 60439;
authors.forename: Julien
authors.initials: J
authors.lastname: Herrou
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125;
authors.forename: Ariane
authors.initials: A
authors.lastname: Briegel
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637;
authors.forename: Grant
authors.initials: G
authors.lastname: Rotskoff
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; Howard Taylor Ricketts Laboratory, University of Chicago, Argonne National Laboratory, Argonne, IL 60439; The Committee on Microbiology, University of Chicago, Chicago, IL 60637 scrosson@uchicago.edu.
authors.forename: Sean
authors.initials: S
authors.lastname: Crosson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D002003:Brucella abortus / Q000166:cytology / Q000472:pathogenicity / Q000502:physiology*; D002455:Cell Division; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D010766:Phosphorylation; D015398:Signal Transduction; D014774:Virulence
chemical_list: D001426:Bacterial Proteins
keywords: Brucella abortus; ChpT; CtrA; cell cycle; two-component system