CodY Promotes Sporulation and Enterotoxin Production by Clostridium perfringens Type A Strain SM101

Li, Jihong; Freedman, John C.; Evans, Daniel R.; McClane, Bruce A.

doi:10.1128/iai.00855-16

Cited by 25 publications

(28 citation statements)

References 41 publications

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“…In liquid, just a single gene encoding a transcriptional regulator, abrB, was expressed at higher levels ( Table 1). AbrB is a global transcriptional regulator that has been shown to be involved in the regulation of sporulation [13] and biofilm formation [14] in C. perfringens. It seems likely that differential expression of abrB in liquid may be due to differences in the nutritional state of the cells, since the CodY protein, which responds to nutritional signals, is a regulator of abrB expression in other strains of C. perfringens [13].…”

Section: Resultsmentioning

confidence: 99%

Changes in the expression of genes encoding type IV pili-associated proteins are seen when Clostridium perfringens is grown in liquid or on surfaces

Soncini

Hartman²,

Gallagher

et al. 2020

BMC Genomics

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Background: Clostridium perfringens is a Gram-positive anaerobic pathogen that causes multiple diseases in humans and animals. C. perfringens lack flagella but have type IV pili (TFP) and can glide on agar surfaces. When C. perfringens bacteria are placed on surfaces, they become elongated, flexible and have TFP on their surface, traits not seen in liquid-grown cells. In addition, the main pilin in C. perfringens TFP, PilA2, undergoes differential posttranslational modification when grown in liquid or on plates. To understand the mechanisms underlying these phenotypes, bacteria were grown in three types of liquid media and on agar plates with the same medium to compare gene expression using RNA-Seq. Results: Hundreds of genes were differentially expressed, including transcriptional regulatory protein-encoding genes and genes associated with TFP functions, which were higher on plates than in liquid. Transcript levels of TFP genes reflected the proportion of each protein predicted to reside in a TFP assembly complex. To measure differences in rates of translation, the Escherichia coli reporter gene gusA gene (encoding β-glucuronidase) was inserted into the chromosome downstream of TFP promoters and in-frame with the first gene of the operon. βglucuronidase expression was then measured in cells grown in liquid or on plates. β-glucuronidase activity was proportional to mRNA levels in liquid-grown cells, but not plate-grown cells, suggesting significant levels of posttranscriptional regulation of these TFP-associated genes occurs when cells are grown on surfaces.Conclusions: This study reveals insights into how a non-flagellated pathogenic rod-shaped bacterium senses and responds to growth on surfaces, including inducing transcriptional regulators and activating multiple posttranscriptional regulatory mechanisms associated with TFP functions.

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Section: Resultsmentioning

confidence: 99%

Changes in the expression of genes encoding type IV pili-associated proteins are seen when Clostridium perfringens is grown in liquid or on surfaces

Soncini

Hartman²,

Gallagher

et al. 2020

BMC Genomics

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“…Measurement of GUS activity. Plasmid pJIR750-sigFp-Gus, prepared previously, was utilized as a ␤-glucuronidase (GUS) reporter vector to monitor expression from the promoter of the spoIIA operon, which encodes the SigF alternate sigma factor (34). For this purpose, 0.2-ml aliquots of F4969 or F4969::nanR transformed with the pJIR750-sigFp-Gus vector were incubated overnight in FTG medium at 37°C, followed by inoculation at 0.2 ml into 10 ml of MDS medium and growth at 43°C for 5 h. The OD 600 of each 5-h culture was measured and cells were collected by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

NanR Regulates Sporulation and Enterotoxin Production by Clostridium perfringens Type F Strain F4969

McClane

2018

Infect Immun

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type F strains, which produce enterotoxin (CPE), are a major cause of gastrointestinal infections, including the second most prevalent bacterial foodborne illness and 5 to 10% cases of antibiotic-associated diarrhea. Virulence of type F strains is primarily ascribable to CPE, which is synthesized only during sporulation. Many type F strains also produce NanI sialidase and carry a operon that likely facilitates uptake and metabolism of sialic acid liberated from glycoconjugates by NanI. During vegetative growth of type F strain F4969, NanR can regulate expression of Given their importance for type F disease, the current study investigated whether NanR can also influence sporulation and CPE production when F4969 or isogenic derivatives are cultured in modified Duncan-Strong sporulation (MDS) medium. An isogenic F4969 null mutant displayed much less sporulation and CPE production but more NanI production than wild-type F4969, indicating that NanR positively regulates sporulation and CPE production but represses NanI production in MDS. Results for the mutant also demonstrated that NanR regulates expression of the operon. A double null mutant mirrored the outcome of the null mutant strain but with a stronger inhibition of sporulation and CPE production, even after overnight incubation. Coupled with results using a null mutant, which had no impairment of sporulation or CPE production, NanR appears to carefully modulate the availability of NanI, operon-encoded proteins and sialic acid to provide sufficient nutrients to sustain sporulation and CPE production when F4969 is cultured in MDS medium.

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“…CodY, first identified in Bacillus subtilis (Slack, Serror, Joyce, & Sonenshein, ), is a DNA‐binding protein and a global regulator of metabolism and virulence genes in nearly all low G+C Gram‐positive pathogens, including Bacillus, Clostridium, Staphylococcus, Streptococcus, Enterococcus and Listeria spp. (Brinsmade, ; Colomer‐Winter, Flores‐Mireles, Kundra, Hultgren, & Lemos, ; Daou et al, ; Geiger & Wolz, ; Li, Freedman, Evans, & McClane, ; Mlynek et al, ; Richardson, Somerville, & Sonenshein, ). The DNA‐binding affinity of CodY from B. subtilis and most other species is increased by interaction with two types of ligands, the branched‐chain amino acids (isoleucine, leucine and valine) and GTP (Han et al, ; Richardson et al, ; Sonenshein, ).…”

Section: Introductionmentioning

confidence: 99%

Genome‐wide identification of Listeria monocytogenes CodY‐binding sites

Biswas

Sonenshein

Belitsky

2020

Molecular Microbiology

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CodY is a global transcriptional regulator that controls, directly or indirectly, the expression of dozens of genes and operons in Listeria monocytogenes. We used in vitro DNA affinity purification combined with massively parallel sequencing (IDAP‐Seq) to identify genome‐wide L. monocytogenes chromosomal DNA regions that CodY binds in vitro. The total number of CodY‐binding regions exceeded 2,000, but they varied significantly in their strengths of binding at different CodY concentrations. The 388 strongest CodY‐binding regions were chosen for further analysis. A strand‐specific analysis of the data allowed pinpointing CodY‐binding sites at close to single‐nucleotide resolution. Gel shift and DNase I footprinting assays confirmed the presence and locations of several CodY‐binding sites. Surprisingly, most of the sites were located within genes’ coding regions. The binding site within the beginning of the coding sequence of the prfA gene, which encodes the master regulator of virulence genes, has been previously implicated in regulation of prfA, but this site was weaker in vitro than hundreds of other sites. The L. monocytogenes CodY protein was functionally similar to Bacillus subtilis CodY when expressed in B. subtilis cells. Based on the sequences of the CodY‐binding sites, a model of CodY interaction with DNA is proposed.

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CodY Promotes Sporulation and Enterotoxin Production by Clostridium perfringens Type A Strain SM101

Cited by 25 publications

References 41 publications

Changes in the expression of genes encoding type IV pili-associated proteins are seen when Clostridium perfringens is grown in liquid or on surfaces

Changes in the expression of genes encoding type IV pili-associated proteins are seen when Clostridium perfringens is grown in liquid or on surfaces

NanR Regulates Sporulation and Enterotoxin Production by Clostridium perfringens Type F Strain F4969

Genome‐wide identification of Listeria monocytogenes CodY‐binding sites

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