An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria

ABSTRACT Signaling by serine/threonine phosphorylation controls diverse processes in bacteria, and identification of the stimuli that activate protein kinases is an outstanding question in the field. Recently, we showed that nutrients stimulate phosphorylation of the protein kinase G substrate GarA...

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Autores principales:	Nabanita Bhattacharyya, Irene Nailain Nkumama, Zaccheus Newland-Smith, Li-Ying Lin, Wen Yin, Rebecca E. Cullen, Jack S. Griffiths, Alexander R. Jarvis, Michael J. Price, Pei Ying Chong, Russell Wallis, Helen M. O’Hare
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Actinobacteria Corynebacterium Mycobacterium tuberculosis sensory transduction processes serine/threonine kinases structural biology Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/19f9a48e73fe41d480209ed39dd2a6a7
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spelling	oai:doaj.org-article:19f9a48e73fe41d480209ed39dd2a6a72021-11-15T16:00:14ZAn Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria10.1128/mBio.00931-182150-7511https://doaj.org/article/19f9a48e73fe41d480209ed39dd2a6a72018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00931-18https://doaj.org/toc/2150-7511ABSTRACT Signaling by serine/threonine phosphorylation controls diverse processes in bacteria, and identification of the stimuli that activate protein kinases is an outstanding question in the field. Recently, we showed that nutrients stimulate phosphorylation of the protein kinase G substrate GarA in Mycobacterium smegmatis and Mycobacterium tuberculosis and that the action of GarA in regulating central metabolism depends upon whether it is phosphorylated. Here we present an investigation into the mechanism by which nutrients activate PknG. Two unknown genes were identified as co-conserved and co-expressed with PknG: their products were a putative lipoprotein, GlnH, and putative transmembrane protein, GlnX. Using a genetic approach, we showed that the membrane protein GlnX is functionally linked to PknG. Furthermore, we determined that the ligand specificity of GlnH matches the amino acids that stimulate GarA phosphorylation. We determined the structure of GlnH in complex with different amino acid ligands (aspartate, glutamate, and asparagine), revealing the structural basis of ligand specificity. We propose that the amino acid concentration in the periplasm is sensed by GlnH and that protein-protein interaction allows transmission of this information across the membrane via GlnX to activate PknG. This sensory system would allow regulation of nutrient utilization in response to changes in nutrient availability. The sensor, signaling, and effector proteins are conserved throughout the Actinobacteria, including the important human pathogen Mycobacterium tuberculosis, industrial amino acid producer Corynebacterium glutamicum, and antibiotic-producing Streptomyces species. IMPORTANCE Tuberculosis (TB) kills 5,000 people every day, and the prevalence of multidrug-resistant TB is increasing in every country. The processes by which the pathogen Mycobacterium tuberculosis senses and responds to changes in its environment are attractive targets for drug development. Bacterial metabolism differs dramatically between growing and dormant cells, and these changes are known to be important in pathogenesis of TB. Here, we used genetic and biochemical approaches to identify proteins that allow M. tuberculosis to detect amino acids in its surroundings so that it can regulate its metabolism. We have also shown how individual amino acids are recognized. The findings have broader significance for other actinobacterial pathogens, such as nontuberculous mycobacteria, as well as Actinobacteria used to produce billions of dollars of amino acids and antibiotics every year.Nabanita BhattacharyyaIrene Nailain NkumamaZaccheus Newland-SmithLi-Ying LinWen YinRebecca E. CullenJack S. GriffithsAlexander R. JarvisMichael J. PricePei Ying ChongRussell WallisHelen M. O’HareAmerican Society for MicrobiologyarticleActinobacteriaCorynebacteriumMycobacterium tuberculosissensory transduction processesserine/threonine kinasesstructural biologyMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Actinobacteria Corynebacterium Mycobacterium tuberculosis sensory transduction processes serine/threonine kinases structural biology Microbiology QR1-502
spellingShingle	Actinobacteria Corynebacterium Mycobacterium tuberculosis sensory transduction processes serine/threonine kinases structural biology Microbiology QR1-502 Nabanita Bhattacharyya Irene Nailain Nkumama Zaccheus Newland-Smith Li-Ying Lin Wen Yin Rebecca E. Cullen Jack S. Griffiths Alexander R. Jarvis Michael J. Price Pei Ying Chong Russell Wallis Helen M. O’Hare An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
description	ABSTRACT Signaling by serine/threonine phosphorylation controls diverse processes in bacteria, and identification of the stimuli that activate protein kinases is an outstanding question in the field. Recently, we showed that nutrients stimulate phosphorylation of the protein kinase G substrate GarA in Mycobacterium smegmatis and Mycobacterium tuberculosis and that the action of GarA in regulating central metabolism depends upon whether it is phosphorylated. Here we present an investigation into the mechanism by which nutrients activate PknG. Two unknown genes were identified as co-conserved and co-expressed with PknG: their products were a putative lipoprotein, GlnH, and putative transmembrane protein, GlnX. Using a genetic approach, we showed that the membrane protein GlnX is functionally linked to PknG. Furthermore, we determined that the ligand specificity of GlnH matches the amino acids that stimulate GarA phosphorylation. We determined the structure of GlnH in complex with different amino acid ligands (aspartate, glutamate, and asparagine), revealing the structural basis of ligand specificity. We propose that the amino acid concentration in the periplasm is sensed by GlnH and that protein-protein interaction allows transmission of this information across the membrane via GlnX to activate PknG. This sensory system would allow regulation of nutrient utilization in response to changes in nutrient availability. The sensor, signaling, and effector proteins are conserved throughout the Actinobacteria, including the important human pathogen Mycobacterium tuberculosis, industrial amino acid producer Corynebacterium glutamicum, and antibiotic-producing Streptomyces species. IMPORTANCE Tuberculosis (TB) kills 5,000 people every day, and the prevalence of multidrug-resistant TB is increasing in every country. The processes by which the pathogen Mycobacterium tuberculosis senses and responds to changes in its environment are attractive targets for drug development. Bacterial metabolism differs dramatically between growing and dormant cells, and these changes are known to be important in pathogenesis of TB. Here, we used genetic and biochemical approaches to identify proteins that allow M. tuberculosis to detect amino acids in its surroundings so that it can regulate its metabolism. We have also shown how individual amino acids are recognized. The findings have broader significance for other actinobacterial pathogens, such as nontuberculous mycobacteria, as well as Actinobacteria used to produce billions of dollars of amino acids and antibiotics every year.
format	article
author	Nabanita Bhattacharyya Irene Nailain Nkumama Zaccheus Newland-Smith Li-Ying Lin Wen Yin Rebecca E. Cullen Jack S. Griffiths Alexander R. Jarvis Michael J. Price Pei Ying Chong Russell Wallis Helen M. O’Hare
author_facet	Nabanita Bhattacharyya Irene Nailain Nkumama Zaccheus Newland-Smith Li-Ying Lin Wen Yin Rebecca E. Cullen Jack S. Griffiths Alexander R. Jarvis Michael J. Price Pei Ying Chong Russell Wallis Helen M. O’Hare
author_sort	Nabanita Bhattacharyya
title	An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
title_short	An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
title_full	An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
title_fullStr	An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
title_full_unstemmed	An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria
title_sort	aspartate-specific solute-binding protein regulates protein kinase g activity to control glutamate metabolism in mycobacteria
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/19f9a48e73fe41d480209ed39dd2a6a7
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An Aspartate-Specific Solute-Binding Protein Regulates Protein Kinase G Activity To Control Glutamate Metabolism in Mycobacteria

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