Título : New substrates and interactors of the mycobacterial Serine/Threonine protein kinase PknG identified by a tailored interactomic approach
Autor(es) : Gil, Magdalena
Lima, Analía
Rivera, Bernardina
Rossello, Jessica
Urdániz, Estefanía
Cascioferro, Alessandro
Carrión, Federico
Wehenkel, Annemarie
Bellinzoni, Marco
Batthyány, Carlos
Pritsch, Otto
Denicola, Ana
Alvarez, María N.
Carvalho, Paulo C.
Lisa, María-Natalia
Brosch, Roland
Piuri, Mariana
Alzari, Pedro M.
Durán, Rosario
Fecha de publicación : mar-2019
Tipo de publicación: Artículo
Versión: Publicado
Publicado por: Elsevier
Publicado en: Journal of Proteomics
Areas del conocimiento : Ciencias Naturales y Exactas
Ciencias Biológicas
Otros descriptores : Proteómica
Ser/Thr quinasas de proteínas
Mycobacterium tuberculosis
Resumen : PknG from Mycobacterium tuberculosis is a multidomain Serine/Threonine protein kinase that regulates bacterial metabolism as well as the pathogen’s ability to survive inside the host by still uncertain mechanisms. To uncover PknG interactome we developed an affinity purification-mass spectrometry strategy to stepwise recover PknG substrates and interactors; and to identify those involving PknG autophosphorylated docking sites. We report a confident list of 7 new putative substrates and 66 direct or indirect partners indicating that PknG regulates many physiological processes, such as nitrogen and energy metabolism, cell wall synthesis and protein translation. GarA and the 50S ribosomal protein L13, two previously reported substrates of PknG, were recovered in our interactome. Comparative proteome analyses of wild type and pknG null mutant M. tuberculosis strains provided evidence that two kinase interactors, the FHA-domain containing protein GarA and the enzyme glutamine synthetase, are indeed endogenous substrates of PknG, stressing the role of this kinase in the regulation of nitrogen metabolism. Interestingly, a second FHA protein was identified as a PknG substrate. Our results show that PknG phosphorylates specific residues in both glutamine synthetase and FhaA in vitro, and suggest that these proteins are phosphorylated by PknG in living mycobacteria.
URI / Handle: https://hdl.handle.net/20.500.12381/3556
DOI: https://doi.org/10.1016/j.jprot.2018.09.013
Institución responsable del proyecto: Instituto Pasteur de Montevideo
Instituto de Investigaciones Biológicas Clemente Estable
Financiadores: Agencia Nacional de Investigación e Innovación
Identificador ANII: FCE_1_2014_1_104045
Nivel de Acceso: Acceso abierto
Licencia CC: Reconocimiento 4.0 Internacional. (CC BY)
Aparece en las colecciones: Institut Pasteur de Montevideo

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