Project description:Blue light (BL) is a major environmental factor that affects the physiology, behavior, and infectivity of bacteria as it contributes to the generation of reactive oxygen species (ROS) while increasing photo-oxidative stress in cells. However, precise photo-oxidative response mechanism in non-phototrophic bacteria is yet to be elucidated. In this study, we investigated the effect of BL in Vibrio cholerae by using genetics and transcriptome profiling. Genome-wide analysis revealed that transcription of 6.3% of V. cholerae genes were regulated by BL. We further showed that BL enhances ROS production, which is generated through the oxidative phosphorylation. To understand signaling mechanisms, we generated several knockouts and analyzed their transcriptome under BL exposure. Studies with a double-knockout confirm an anti-sigma factor (ChrR) and putative metalloregulatory-like protein (MerR) are responsible for the genome-wide regulation to BL response in V. cholerae. Collectively, these results demonstrate that MerR-like proteins, in addition to ChrR, are required for V. cholerae to mount an appropriate response against photo-oxidative stress induced by BL. Outside its natural host, V. cholerae can survive for extended periods in natural aquatic environments. Therefore, the regulation of light response for V. cholerae may be a critical cellular process for its survival in these environments.

2017-01-26 | GSE79911 | GEO

Comparative whole cell proteomics data of Vibrio cholerae O1 biovar El Tor str. N16961 versus full length cgtA knockdown and CgtA(∆CTD) during nutrient limiting condition

Project description:Adapting to nutritional downshifts is crucial for the survival of Vibrio cholerae. CgtA, a 50S ribosome-associated essential GTPase, is a bonafide stringent response protein. CgtA, in association with SpoT, modulates the intracellular (p)ppGpp alarmone levels in a nutrient-rich environment. We studied the influence of CgtA during the growth of V. cholerae in a minimal medium in contrast to its pre-defined role in a nutrient-rich medium. Here, we show the pleiotropic effects of CgtA on growth, viability, motility, morphology, and persister phenotype of a V. cholerae strain where the full-length wildtype (Wt) cgtA was deleted. An in-frame deletion of the 52 amino acids long unstructured C-terminal domain of the CgtA GTPase defined its in vivo functionality. Proteomic analyses revealed that loss of CgtA significantly altered 311 proteins involved in diverse cellular processes. However, the CgtA C-terminus deleted strain altered 240 proteins with a major overlap with the full-length cgtA deleted condition. A sustained mRNA expression pattern of CgtA is observed in a minimal medium. Whereas, in nutrient-rich LB medium, intermittent expression of cgtA is observed with the highest expression during the late-logarithmic to early-stationary phase. We propose that minimal media-associated nutrient stress coupled to cgtA depletion aggravates the intracellular stress in V. cholerae, leading to abnormal protein synthesis, altered DNA replication, and transcriptional machinery, negatively affecting cellular energy metabolism and many vital processes. This study suggests that the nutrient-media-dependent controlled expression of CgtA is a mechanism by which V. cholerae can remodel its transcriptome and proteome. Our study reveals an alternative facet of the survival of V. cholerae during nutritional downshift and the consequences concomitantly with cgtA knockdown as evident from the complex altered regulatory network.

2023-03-03 | PXD034015 | Pride

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