Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

The Signal Transduction Protein PII Controls the Levels of the Cyanobacterial Multitasking Protein PipX

Version 1 : Received: 6 September 2023 / Approved: 7 September 2023 / Online: 7 September 2023 (11:23:14 CEST)

A peer-reviewed article of this Preprint also exists.

Llop, A.; Tremiño, L.; Cantos, R.; Contreras, A. The Signal Transduction Protein PII Controls the Levels of the Cyanobacterial Protein PipX. Microorganisms 2023, 11, 2379. Llop, A.; Tremiño, L.; Cantos, R.; Contreras, A. The Signal Transduction Protein PII Controls the Levels of the Cyanobacterial Protein PipX. Microorganisms 2023, 11, 2379.

Abstract

Cyanobacteria, microorganisms performing oxygenic photosynthesis, must adapt their metabolic processes to environmental challenges such as day and night changes. PipX, a unique regulatory protein from cyanobacteria provides a mechanistic link between signalling protein PII, a widely conserved (in bacteria and plants) transducer of carbon/nitrogen/energy richness, and the transcriptional regulator NtcA, which controls a large regulon involved in nitrogen assimilation. PipX is also involved in translational regulation by interaction with the ribosome-assembly GTPase EngA. However, increases of the PipX/PII ratio are toxic, presumably by abnormally increased binding of PipX to other partner(s). Here we present mutational and structural analyses of reported PipX-PII and PipX-NtcA complexes, leading to identification of single amino acid changes that decrease or abolish PipX toxicity. Four out of 11 mutations decreasing toxicity did not decrease PipX levels, suggesting that the targeted residues (F12, D23, L36, R54) provide toxicity determinants. In addition, one of those four mutations (D23A) argued against the over-activation of NtcA as the cause of PipX toxicity. Since most mutations at residues contacting PII decreased PipX levels, PipX stability would depend on its ability to bind to PII, a conclusion supported by the light-induced decrease in PipX levels observed in Synechococcus elongatus PCC7942.

Keywords

NtcA; EngA; Synechococcus elongatus; nitrogen regulation network; light and dark conditions; PipX toxicity; energy sensing; mutational analysis

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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