title: The extracytoplasmic domain of the Mycobacterium tuberculosis Ser/Thr kinase PknB binds specific muropeptides and is required for PknB localization.
abstract: The Mycobacterium tuberculosis Ser/Thr kinase PknB has been implicated in the regulation of cell growth and morphology in this organism. The extracytoplasmic domain of this membrane protein comprises four penicillin binding protein and Ser/Thr kinase associated (PASTA) domains, which are predicted to bind stem peptides of peptidoglycan. Using a comprehensive library of synthetic muropeptides, we demonstrate that the extracytoplasmic domain of PknB binds muropeptides in a manner dependent on the presence of specific amino acids at the second and third positions of the stem peptide, and on the presence of the sugar moiety N-acetylmuramic acid linked to the peptide. We further show that PknB localizes strongly to the mid-cell and also to the cell poles, and that the extracytoplasmic domain is required for PknB localization. In contrast to strong growth stimulation by conditioned medium, we observe no growth stimulation of M. tuberculosis by a synthetic muropeptide with high affinity for the PknB PASTAs. We do find a moderate effect of a high affinity peptide on resuscitation of dormant cells. While the PASTA domains of PknB may play a role in stimulating growth by binding exogenous peptidoglycan fragments, our data indicate that a major function of these domains is for proper PknB localization, likely through binding of peptidoglycan fragments produced locally at the mid-cell and the cell poles. These data suggest a model in which PknB is targeted to the sites of peptidoglycan turnover to regulate cell growth and cell division.
authors.affiliation: Division of Infectious Diseases, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Mushtaq
authors.initials: M
authors.lastname: Mir
authors.forename: Jinkeng
authors.initials: J
authors.lastname: Asong
authors.forename: Xiuru
authors.initials: X
authors.lastname: Li
authors.forename: Jessica
authors.initials: J
authors.lastname: Cardot
authors.forename: Geert-Jan
authors.initials: GJ
authors.lastname: Boons
authors.forename: Robert N
authors.initials: RN
authors.lastname: Husson
mesh_terms: D002455:Cell Division / Q000502:physiology*; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D010455:Peptides; D010457:Peptidoglycan; C475060:PknB protein, Mycobacterium tuberculosis; D017346:Protein Serine-Threonine Kinases