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Investigating the role of the carbon storage regulator A (CsrA) in Leptospira spp.

Abstract : Carbon Storage Regulator A (CsrA) is a well-characterized post-transcriptional global regulator that plays a critical role in response to environmental changes in many bacteria. CsrA has been reported to regulate several metabolic pathways, motility, biofilm formation, and virulence-associated genes. The role of csrA in Leptospira spp., which are able to survive in different environmental niches and infect a wide variety of reservoir hosts, has not been characterized. To investigate the role of csrA as a gene regulator in Leptospira , we generated a L . biflexa csrA deletion mutant (Δ csrA ) and csrA overexpressing Leptospira strains. The Δ csrA L . biflexa displayed poor growth under starvation conditions. RNA sequencing revealed that in rich medium only a few genes, including the gene encoding the flagellar filament protein FlaB3, were differentially expressed in the Δ csrA mutant. In contrast, 575 transcripts were differentially expressed when csrA was overexpressed in L . biflexa . Electrophoretic mobility shift assay (EMSA) confirmed the RNA-seq data in the Δ csrA mutant, showing direct binding of recombinant CsrA to flaB3 mRNA. In the pathogen L . interrogans , we were not able to generate a csrA mutant. We therefore decided to overexpress csrA in L . interrogans . In contrast to the overexpressing strain of L . biflexa , the overexpressing L . interrogans strain had poor motility on soft agar. The overexpressing strain of L . interrogans also showed significant upregulation of the flagellin flaB1 , flaB2 , and flaB4 . The interaction of L . interrogans rCsrA and flaB4 was confirmed by EMSA. Our results demonstrated that CsrA may function as a global regulator in Leptospira spp. under certain conditions that cause csrA overexpression. Interestingly, the mechanisms of action and gene targets of CsrA may be different between non-pathogenic and pathogenic Leptospira strains.
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https://hal-pasteur.archives-ouvertes.fr/pasteur-03509058
Contributor : Thomas Cokelaer Connect in order to contact the contributor
Submitted on : Monday, January 3, 2022 - 11:39:42 PM
Last modification on : Friday, August 5, 2022 - 12:03:02 PM
Long-term archiving on: : Monday, April 4, 2022 - 9:26:11 PM

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Theerapat Phoka, Lenka Fule, Juliana Pipoli da Fonseca, Thomas Cokelaer, Mathieu Picardeau, et al.. Investigating the role of the carbon storage regulator A (CsrA) in Leptospira spp.. PLoS ONE, 2021, 16 (12), pp.e0260981. ⟨10.1371/journal.pone.0260981⟩. ⟨pasteur-03509058⟩

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