Dataset Information

Proteomic and metabolomic analysis of Azospirillum brasilense ntrC mutant under high and low nitrogen conditions

ABSTRACT: Azospirillum brasilense is a free-living diazothophic bacterium that associates with important grasses and cereals, promoting plant growth and increasing crop productivity. The nitrogen metabolism in this organism is tightly regulated by the availability of nitrogen and by the Ntr regulatory system. This system comprises the GlnD protein, which is capable of adding uridylyl groups to the PII proteins, GlnB and GlnZ, under limiting nitrogen levels. Under such conditions, the histidine kinase NtrB cannot interact with GlnB and phosphorylate NtrC. When phosphorylated, NtrC is a transcriptional activator of genes involved in metabolism of alternative nitrogen sources. Considering the key role of NtrC in nitrogen metabolism in Azospirillum brasilense, in this work we evaluated the proteomic and metabolomic profiles of the wild type FP2 strain and its mutant ntrC grown under high and low nitrogen. Analysis of the integrated data identifies, novel NtrC targets, including proteins involved in the response against oxidative stressnm (i.e., glutathione S-transferase and hydroperoxide resistance protein,), underlining the importance of NtrC to bacterial survival under oxidative stress conditions.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Azospirillum Brasilense Fp2

SUBMITTER: Gustavo De Souza

LAB HEAD: Emmanuel M Souza

PROVIDER: PXD014120 | Pride | 2019-10-14

REPOSITORIES: Pride

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Proteomic and Metabolomic Analysis of Azospirillum brasilensentrC Mutant under High and Low Nitrogen Conditions.

Kukolj Caroline C Pedrosa Fábio O FO de Souza Gustavo A GA Sumner Lloyd W LW Lei Zhentian Z Sumner Barbara B do Amaral Fernanda P FP Juexin Wang W Trupti Joshi J Huergo Luciano F LF Monteiro Rose Adele RA Valdameri Glaucio G Stacey Gary G de Souza Emanuel M EM

Journal of proteome research 20191025 1

Azospirillum brasilense is a diazotrophic microorganism capable of associating with roots of important grasses and cereals, promoting plant growth and increasing crop yields. Nitrogen levels and the Ntr regulatory system control the nitrogen metabolism in A. brasilense. This system comprises the nitrogen regulatory proteins GlnD, which is capable of adding uridylyl groups to the PII proteins, GlnB (PII-1) and GlnZ (PII-2), under limiting nitrogen levels. Under such conditions, the ...[more]

PMID: 31599156

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Project description:Caulobacter species are dimorphic Gram-negative bacteria that inhabit diverse terrestrial and aquatic ecosystems. A suite of molecular sensory systems enables these microbes to control growth, development, and reproduction in response to levels of essential elements, including carbon, nitrogen, and phosphorus. Although the enhancer binding protein NtrC and its cognate sensor histidine kinase NtrB are well-established regulators of bacterial nitrogen assimilation, their precise functions in Caulobacter metabolism and cell development remained largely undefined. Deletion of C. crescentus ntrC slowed cell growth in complex medium, while ntrB and ntrC were essential for growth when ammonium was the sole nitrogen source due to their requirement for glutamine synthase (glnA) expression. Random transposition of a conserved IS3-family mobile genetic element frequently rescued the growth defect of ntrC mutant strains by restoring glnBA transcription, revealing a possible role for IS3 transposition in shaping the evolution of Caulobacter populations during nitrogen limitation. We further defined the NtrC regulon through transcriptomic and ChIP-seq analyses, which demonstrated the role of NtrC as both a transcriptional activator and repressor. The chromosome of C. crescentus harbors dozens of direct binding sites for NtrC, with a significant proportion located near genes involved in polysaccharide biosynthesis. Numerous NtrC binding sites align with those of GapR, an essential protein involved in chromosome organization, and MucR1, a cell cycle regulator. This observation suggests that NtrC participates in the regulation of cell cycle and cell development. Indeed, loss of NtrC function led to elongated polar stalks and elevated synthesis of cell envelope polysaccharides. These developmental defects were restored by supplementing media with glutamine or by ectopic expression of the glnBA operon. This study establishes a regulatory connection between NtrC, nitrogen metabolism, polar morphogenesis, and envelope polysaccharide synthesis in Caulobacter.

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Dataset Information

Proteomic and metabolomic analysis of Azospirillum brasilense ntrC mutant under high and low nitrogen conditions

Dataset's files

Publications

Proteomic and Metabolomic Analysis of <i>Azospirillum brasilense</i><i>ntrC</i> Mutant under High and Low Nitrogen Conditions.

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