Dataset Information

The external PASTA domain of the essential serine/threonine protein kinase PknB regulates mycobacterial growth

ABSTRACT: PknB is an essential serine/threonine protein kinase required for mycobacterial cell division and cell wall biosynthesis. Here we demonstrate that over-expression of the external PknB_PASTA domain in mycobacteria results in delayed regrowth, accumulation of elongated bacteria and increased sensitivity to -lactam antibiotics. These changes are accompanied by altered production of certain enzymes involved in biosynthesis of cell wall as revealed by proteomics studies. The growth inhibition caused by over-expression of the PknB_PASTA domain is completely abolished by enhanced concentration of magnesium ions but not muropeptides. Finally, we show that the addition of recombinant PASTA domain could prevent regrowth of Mycobacterium tuberculosis and therefore offers an alternative opportunity to control replication of this pathogen. These results suggest that the PknB_PASTA domain is involved in regulation of peptidoglycan biosynthesis and maintenance of cell wall architecture.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mycobacterium Smegmatis

TISSUE(S): Cell Culture

SUBMITTER: Andrew Bottrill

LAB HEAD: Galina V. Mukamolova

PROVIDER: PXD002120 | Pride | 2018-03-13

REPOSITORIES: Pride

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Publications

The external PASTA domain of the essential serine/threonine protein kinase PknB regulates mycobacterial growth.

Turapov Obolbek O Loraine Jessica J Jenkins Christopher H CH Barthe Philippe P McFeely Daniel D Forti Francesca F Ghisotti Daniela D Hesek Dusan D Lee Mijoon M Bottrill Andrew R AR Vollmer Waldemar W Mobashery Shahriar S Cohen-Gonsaud Martin M Mukamolova Galina V GV

Open biology 20150701 7

PknB is an essential serine/threonine protein kinase required for mycobacterial cell division and cell-wall biosynthesis. Here we demonstrate that overexpression of the external PknB_PASTA domain in mycobacteria results in delayed regrowth, accumulation of elongated bacteria and increased sensitivity to β-lactam antibiotics. These changes are accompanied by altered production of certain enzymes involved in cell-wall biosynthesis as revealed by proteomics studies. The growth inhibition caused by ...[more]

PMID: 26136255

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Project description:Stem internodes of grasses function in mechanical support, transport, and, in some species are a major sink organ for carbon in the form of cell wall polymers. To establish the rice elongating internode as a model for secondary cell wall development, we conducted cell wall composition, proteomic and metabolomic analyses of the second rice internode at booting stage. We measured major secondary cell wall components along eight segments of an elongating internode. Cellulose, lignin, and xylose increase as a percentage of cell wall material from the younger to the older internode segments, indicating active cell wall synthesis. With the whole elongating internode, we measured peptides via liquid-chromatography mass spectrometry (LC-MS) following trypsin digestion of size fractionated proteins. This identified a total of 3249 protein groups with at least two unique peptides, including many glycosyltransferases, acyltransferases, glycohyrolases, cell wall-localized proteins, and protein kinases that have or may have functions in cell wall biosynthesis or remodeling. In addition, GO over-representation analysis and KO pathway analysis indicate many proteins involved in biosynthetic processes, especially the synthesis of secondary metabolites such as phenylpropanoids, flavonoids, and tepenoids. Therefore, we also used LC-MS to measure methanol-extracted secondary metabolites from the whole internode at the elongation stage and three post-elongation stages, and from leaf and root at the second post-elongation stage. The results indicate secondary metabolites in stems are distinct from those of roots and leaves, and show different profiles during stem development. This study fills a void of knowledge of proteomics and metabolomics data for grass stems, especially of rice, and provides baseline knowledge for more detailed studies of cell wall synthesis and other biological processes during internode development. This and future work is aimed at optimizing stem development and cell wall composition of grasses to improve agronomic properties and biofuel production.

Dataset Information

The external PASTA domain of the essential serine/threonine protein kinase PknB regulates mycobacterial growth

Publications

The external PASTA domain of the essential serine/threonine protein kinase PknB regulates mycobacterial growth.

Similar Datasets

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