The Role of Regulator Catabolite Control Protein A (CcpA) in Streptococcus agalactiae Physiology and Stress Response

Roux, Anne-Emmanuelle; Robert, Sylvie; Bastat, Mathilde; Rosinski‐Chupin, Isabelle; Rong, Vanessa; Holbert, Sébastien; Mereghetti, Laurent; Camiade, Émilie

doi:10.1128/spectrum.02080-22

Cited by 4 publications

(2 citation statements)

References 91 publications

(117 reference statements)

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“…A separate Tn-seq investigation of GBS strain CJB111 found ccpA to be essential in chemically defined media, but not Todd Hewitt broth 56 . This finding was reconfirmed by another group, which was able to generate a ΔccpA deletion mutant in A909 grown in Todd Hewitt broth 68 . We also targeted the gyrA gene, which encodes the gyrase A subunit of the DNA gyrase enzyme, which is essential for bacterial viability 54,69,70 .…”

Section: Crispri Of Gbs Essential Gene Targetsmentioning

confidence: 53%

Group BStreptococcusCas9 variants provide insight into programmable gene repression and CRISPR-Cas transcriptional effects

Gopalakrishna

Hillebrand

Bhavana

et al. 2023

Preprint

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Group B Streptococcus (GBS; S. agalactiae) causes chorioamnionitis, neonatal sepsis,and can also cause disease in healthy or immunocompromised adults. GBS possesses a type II-A CRISPR-Cas9 system, which defends against foreign DNA within the bacterial cell. Several recent publications have shown that GBS Cas9 influences genome-wide transcription through a mechanism uncoupled from its function as a specific, RNA-programmable endonuclease. We examine GBS Cas9 effects on genome-wide transcription through generation of several isogenic variants with specific functional defects. We compare whole-genome RNA-seq from Δcas9 GBS with a full-length Cas9 gene deletion; dcas9 defective in its ability to cleave DNA but still able to bind to frequently occurring protospacer adjacent motifs; and scas9 that retains its catalytic domains but is unable to bind protospacer adjacent motifs. Comparing scas9 GBS to the other variants, we identify nonspecific protospacer adjacent motif binding as a driver of genome-wide, Cas9 transcriptional effects in GBS. We also show that Cas9 transcriptional effects from nonspecific scanning tend to influence genes involved in bacterial defense and nucleotide or carbohydrate transport and metabolism. While genome-wide transcription effects are detectable by analysis of next-generation sequencing, they do not result in virulence changes in a mouse model of sepsis. We also demonstrate that catalytically inactive dCas9 expressed from the GBS chromosome can be used with a straightforward, plasmid-based, single guide RNA expression system to suppress transcription of specific GBS genes without potentially confounding off-target effects. We anticipate that this system will be useful for study of nonessential and essential gene roles in GBS physiology and pathogenesis.

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Section: Crispri Of Gbs Essential Gene Targetsmentioning

confidence: 53%

Group BStreptococcusCas9 variants provide insight into programmable gene repression and CRISPR-Cas transcriptional effects

Gopalakrishna

Hillebrand

Bhavana

et al. 2023

Preprint

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“…This gene has been reported to be essential for S. agalactiae survival [ 40 ]. However, Roux et al [ 41 ] deleted the ccpA gene in S. agalactiae strain A909 successfully. No more than 95% query cover exist between the genomes of the piscine GD201008-001 and human-derived isolate A909, which might explain the contradiction between different researches.…”

Section: Discussionmentioning

confidence: 99%

luxS contributes to intramacrophage survival of Streptococcus agalactiae by positively affecting the expression of fruRKI operon

Cao,

Dong,

Guo

et al. 2023

Vet Res

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The LuxS quorum sensing system is a widespread system employed by many bacteria for cell-to-cell communication. The luxS gene has been demonstrated to play a crucial role in intramacrophage survival of piscine Streptococcus agalactiae, but the underlying mechanism remains largely unknown. In this study, transcriptome analysis, followed by the luxS gene deletion and subsequent functional studies, confirmed that impaired bacterial survival inside macrophages due to the inactivation of luxS was associated with reduced transcription of the fruRKI operon, encoding the fructose-specific phosphotransferase system. Further, luxS was determined not to enhance the transcription of fruRKI operon by binding its promoter, but to upregulate the expression of this operon via affecting the binding ability of catabolite control protein A (CcpA) to the catabolite responsive element (cre) in the promoter of fruRKI. Collectively, our study identifies a novel and previously unappreciated role for luxS in bacterial intracellular survival, which may give a more thorough understanding of the immune evasion mechanism in S. agalactiae.

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Regulation of PhoB on biofilm formation and hemolysin gene hlyA and ciaR of Streptococcus agalactiae

Cai,

Yang,

Peng

et al. 2024

Veterinary Microbiology

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The Role of Regulator Catabolite Control Protein A (CcpA) in Streptococcus agalactiae Physiology and Stress Response

Cited by 4 publications

References 91 publications

Group BStreptococcusCas9 variants provide insight into programmable gene repression and CRISPR-Cas transcriptional effects

Group BStreptococcusCas9 variants provide insight into programmable gene repression and CRISPR-Cas transcriptional effects

luxS contributes to intramacrophage survival of Streptococcus agalactiae by positively affecting the expression of fruRKI operon

Regulation of PhoB on biofilm formation and hemolysin gene hlyA and ciaR of Streptococcus agalactiae

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