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

Abstract Estrogens originating from human and animal excretion, as well as from anthropogenic sources such as cosmetics, plastics, pesticides, detergents, and pharmaceuticals, are among the most concerning endocrine-disrupting compounds in aquatic environments due to their potent estrogenic activity. While their effects on fish reproduction are well documented, their impact on development, particularly metamorphosis, remains poorly studied. This hormonal transition, mainly controlled by the thyroid axis, is essential for the shift from the larval to the juvenile stage in teleosts.The effects of two contraceptive estrogens on zebrafish (Danio rerio) metamorphosis were evaluated: 17α-ethinylestradiol (EE2), a synthetic reference estrogen, and estetrol (E4), a natural estrogen recently introduced in a new combined oral contraceptive formulation. Continuous exposure from fertilization to the end of metamorphosis allowed the assessment of morphological changes, disruptions of the thyroid axis, and modifications of additional molecular pathways potentially involved in metamorphic regulation.EE2 induced significant delays and disturbances in metamorphosis, affecting both internal and external morphological traits, confirming its role as an endocrine disruptor of concern. In contrast, E4 did not cause any detectable morphological alterations even at concentrations far exceeding those expected in the environment, indicating a limited ecotoxicological risk. Molecular analyses showed that EE2 strongly affected thyroid signaling and energy metabolism during metamorphosis, whereas E4 induced only minor transcriptional and proteomic changes.This study provides the first evidence that EE2 can disrupt zebrafish metamorphosis and highlights the importance of including this developmental stage in ecotoxicological assessments. The results also suggest a larger environmental safety margin for E4, although further research is needed to clarify the mechanisms linking estrogen exposure to metamorphic regulation.JuryProf. Frederik DE LAENDER (UNamur), PresidentProf. Patrick KESTEMONT (UNamur), SecretaryDr. Sébastien BAEKELANDT (UNamur)Dr. Valérie CORNET (UNamur)Prof. Jean-Baptiste FINI (Muséum National d'Histoire Naturelle, Paris)Dr. Marc MULLER (ULiège)Prof. Veerle DARRAS (KULeuven)