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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Reconocimiento 4.0 Internacional. (CC BY)