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Geyman, L.

Publications and source records attributed to Geyman, L..

3 recordsLinked to original sources

Catabolite Activator Protein and quorum sensing cross-control group behaviors in Vibrio campbellii

Vibrio species adapt to different niches by sensing and responding to environmental signals such as nutrients, host cues, and quorum sensing autoinducers. Vibrio campbellii uses the master quorum sensing transcription factor LuxR to control expression of hundreds of genes at high cell density. Furthermore, many gamma-proteobacteria use the global transcription factor Catabolite Activator Protein (CAP) to control numerous physiologically relevant pathways, many of which are also modulated by quorum sensing, such as competence, biofilm formation, and carbon metabolism. However, the extent to which these two global transcription factors overlap to co-regulate gene expression and bacterial behaviors is understudied. In this work, we used ChIP-seq and RNA-seq to determine the individual and combined regulons of CAP and LuxR in V. campbellii. We found that CAP and LuxR co-occupied 11 promoters to synergistically or antagonistically co-regulate genes involved with metabolism, respiration, and virulence. It was previously proposed that CAP and LuxR both bound the bioluminescence (luxCDABE) promoter to co-regulate these genes, and our RNA-seq data showed that these were indeed the most strongly co-regulated genes. However, our ChIP-seq data revealed that only LuxR bound the luxCDABE promoter in vivo. This pattern of co-regulation--where both CAP and LuxR strongly impact transcription but only LuxR binds the promoter--was the most common mechanism observed. Our model for bioluminescence regulation is that CAP indirectly activates luxCDABE expression through the regulation of an intermediate factor. This study established new connections between the global gene regulatory networks underpinning nutrient sensing and population sensing. ImportanceVibrio bacteria (vibrios) colonize and infect diverse marine hosts including corals, fish, oysters, and shrimp, and can also cause life-threatening human infections, all of which are rising annually due to increasing ocean temperatures. To develop effective treatments against Vibrio infections, it is critical to understand the global gene regulation mechanisms vibrios use that enable pathogenic lifestyles. Signal transduction systems in bacteria are well-characterized; however, how vibrios coordinate global gene expression changes in response to multiple environmental inputs is understudied. Here, we determined how the aquaculture pathogen Vibrio campbellii regulates global gene expression in response to bacterial population signals and nutrient availability signals. Our findings help contextualize how vibrios respond to their fluctuating environments in nature.

microbiology↗

Quorum sensing employs a dual regulatory mechanism to repress T3SS gene expression

The type III secretion system (T3SS) is a needle-like complex used by numerous bacterial pathogens in host infection by directly injecting exotoxins into the host cell cytoplasm, leading to cell death. The T3SS is a known virulence factor in the shrimp pathogen Vibrio campbellii. The [~]40 genes comprising the V. campbellii T3SS are regulated by a network of transcription factors in response to changes in the cells environment: cell density (quorum sensing; QS), temperature, calcium, and host cell contact. Under positive environmental stimuli, the master T3SS transcription factor ExsA activates expression of the four structural T3SS operons required for needle formation. Previous studies identified a key role of the master QS transcription factor LuxR: repression of exsA transcription via DNA binding at the exsBA promoter. Here we uncovered a new regulatory role of LuxR: indirect post-translational repression of ExsA activity via direct transcriptional repression of the gene encoding the anti-anti-activator ExsC. In V. campbellii, ExsC is a positive regulator of T3SS transcription: high ExsC expression leads to full ExsA transcription activation of the T3SS structural promoters. LuxR binding at the exsC promoter represses transcription of exsC through disruption of ExsA binding. Our findings collectively show that V. campbellii responds to high cell density signals to shut down the expression of the T3SS. We postulate that this dual regulatory mechanism by LuxR enables both the rapid inactivation of existing ExsA protein and blocks its further synthesis, leading to a rapid shutdown of T3SS activity at high cell density. ImportanceVibrio campbellii utilizes the type III secretion system (T3SS) as a mechanism of pathogenesis, which is a highly studied injectisome complex that delivers exotoxins into host cells during infection. The T3SS pathogenicity island in V. campbellii comprises [~]40 genes that are organized into four structural operons. In this study, we determined that quorum sensing - a method of bacterial communication - regulates T3SS genes both at the transcriptional and post-translational levels to shut down T3SS gene expression at high population densities.

microbiology↗

Vibrio campbellii DS40M4 is a tractable model strain that diverges from the canonical quorum-sensing regulatory circuit in vibrios

Vibrio campbellii BB120 (previously classified as Vibrio harveyi) is a fundamental model strain for studying quorum sensing in vibrios. A phylogenetic evaluation of sequenced Vibrio strains in Genbank revealed that BB120 is closely related to the environmental isolate V. campbellii DS40M4. We exploited DS40M4s competence for exogenous DNA uptake to rapidly generate >30 isogenic strains with deletions of genes encoding BB120 quorum-sensing system homologs. Our results show that the quorum-sensing circuit of DS40M4 is distinct from BB120 in three ways: 1) DS40M4 does not produce an acyl homoserine lactone autoinducer but encodes an active orphan LuxN receptor, 2) the quorum regulatory small RNAs (Qrrs) are not solely regulated by autoinducer signaling through the response regulator LuxO, and 3) the DS40M4 quorum-sensing regulon is much smaller than BB120 (~100 genes vs ~400 genes, respectively). Using comparative genomics to expand our understanding of quorum-sensing circuit diversity, we observe that conservation of LuxM/LuxN proteins differs widely both between and within Vibrio species. These strains are also phenotypically distinct: DS40M4 exhibits stronger interbacterial cell killing, whereas BB120 forms more robust biofilms and is bioluminescent. These results underscore the need to examine wild isolates for a broader view of bacterial diversity in the marine ecosystem. Originality-Significance StatementWild bacterial isolates yield important information about traits that vary within species. Here, we compare environmental isolate Vibrio campbellii DS40M4 to its close relative, the model strain BB120 that has been a fundamental strain for studying quorum sensing for >30 years. We examine several phenotypes that define this species, including quorum sensing, bioluminescence, and biofilm formation. Importantly, DS40M4 is naturally transformable with exogenous DNA, which allows for the rapid generation of mutants in a laboratory setting. By exploiting natural transformation, we genetically dissected the functions of BB120 quorum-sensing system homologs in the DS40M4 strain, including two-component signaling systems, transcriptional regulators, and small RNAs.

microbiology↗