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NAD+ biosynthesis in bacteria is controlled by global carbon/nitrogen levels via PII signaling.

ABSTRACT: NAD+ is a central metabolite participating in core metabolic redox reactions. The prokaryotic NAD synthetase enzyme NadE catalyzes the last step of NAD+ biosynthesis, converting nicotinic acid adenine dinucleotide (NaAD) to NAD+ Some members of the NadE family use l-glutamine as a nitrogen donor and are named NadEGln Previous gene neighborhood analysis has indicated that the bacterial nadE gene is frequently clustered with the gene encoding the regulatory signal transduction protein PII, suggesting a functional relationship between these proteins in response to the nutritional status and the carbon/nitrogen ratio of the bacterial cell. Here, using affinity chromatography, bioinformatics analyses, NAD synthetase activity, and biolayer interferometry assays, we show that PII and NadEGln physically interact in vitro, that this complex relieves NadEGln negative feedback inhibition by NAD+ This mechanism is conserved in distantly related bacteria. Of note, the PII protein allosteric effector and cellular nitrogen level indicator 2-oxoglutarate (2-OG) inhibited the formation of the PII-NadEGln complex within a physiological range. These results indicate an interplay between the levels of ATP, ADP, 2-OG, PII-sensed glutamine, and NAD+, representing a metabolic hub that may balance the levels of core nitrogen and carbon metabolites. Our findings support the notion that PII proteins act as a dissociable regulatory subunit of NadEGln, thereby enabling the control of NAD+ biosynthesis according to the nutritional status of the bacterial cell.

SUBMITTER: Santos ARS

PROVIDER: S-EPMC7196632 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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NAD+ biosynthesis in bacteria is controlled by global carbon/nitrogen levels via PII signaling.

Santos Adrian Richard Schenberger ARS Gerhardt Edileusa Cristina Marques ECM Parize Erick E Pedrosa Fabio Oliveira FO Steffens Maria Berenice Reynaud MBR Chubatsu Leda Satie LS Souza Emanuel Maltempi EM Passaglia Luciane Maria Pereira LMP Sant'Anna Fernando Hayashi FH de Souza Gustavo Antônio GA Huergo Luciano Fernandes LF Forchhammer Karl K

The Journal of biological chemistry 20200316 18

NAD+ is a central metabolite participating in core metabolic redox reactions. The prokaryotic NAD synthetase enzyme NadE catalyzes the last step of NAD+ biosynthesis, converting nicotinic acid adenine dinucleotide (NaAD) to NAD+ Some members of the NadE family use l-glutamine as a nitrogen donor and are named NadEGln Previous gene neighborhood analysis has indicated that the bacterial nadE gene is frequently clustered with the gene encoding the ...[more]

PMID: 32179648

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NAD+ biosynthesis in bacteria is controlled by global carbon/nitrogen levels via PII signaling.

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NAD<sup>+</sup> biosynthesis in bacteria is controlled by global carbon/nitrogen levels via PII signaling.

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