Soutenance publique de thèse de doctorat en Sciences biologiques - Aishwarya Saxena

Elucidation of the (p)ppGpp-dependent transcriptomic landscape in Caulobacter crescentus

Jury

• Prof. Gipsi LIMA MENDEZ (UNamur), President
• Prof Régis HALLEZ (UNamur), Secretary
• Dr Emanuele BIONDI (CNRS-Université Paris-Saclay)
• Prof. Justine COLLIER (University of Lausanne)
• Dr Marie DELABY (Université de Montréal)

Abstract

Primarily described as an alarmone, secondary messenger (p)ppGpp, when accumulated, binds to many targets involved in DNA replication, translation, and transcription. In the asymmetrically-dividing a-proteobacterium Caulobacter crescentus, (p)ppGpp has been shown to strongly impact cell cycle progression and differentiation, promoting the non-replicating G1/swarmer phase. Mutations in the major subunits of transcriptional complex, b or b’ subunits, were able to display the (p)ppGpp-related phenotypes even in the absence of the alarmone. We identified that the transcriptional holo-enzyme, RNA polymerase (RNAP) is a primary target of (p)ppGpp in C. crescentus. Furthermore, mutations that inactivate (p)ppGpp binding to RNAP annihilated the (p)ppGpp-related phenotypes and phenocopied a (p)ppGpp⁰ strain. Our RNAseq analysis further elucidated the changes in the transcriptional landscape of C. crescentus cells displaying different (p)ppGpp levels and expressing RNAP mutants. Since the DNA replication initiation protein DnaA is required to exit the G1 phase, we observed that it was significantly less abundant in cells accumulating (p)ppGpp. We further explored its proteolysis under the influence of (p)ppGpp. Our work suggests that (p)ppGpp regulates cell cycle and differentiation in C. crescentus by reprogramming transcription and triggering proteolytic degradation of key cell cycle regulators by yet unknown mechanisms. In Part II, we identified two σ factors belonging to the ECF family that might be involved in this (p)ppGpp-accompanied phenotypes. In Part III, we propose an overlapping role of the ω subunit, RpoZ, and the heat shock subunit, RpoH, in carbon metabolism.

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