Contributions of NanI Sialidase to Caco-2 Cell Adherence by Clostridium perfringens Type A and C Strains Causing Human Intestinal Disease

Li, Jihong; McClane, Bruce A.

doi:10.1128/iai.02322-14

Cited by 41 publications

(125 citation statements)

References 43 publications

Supporting

Mentioning

113

Contrasting

Order By: Relevance

“…All qRT-PCR primers were designed using the Integrated DNA Technologies (IDT) website. The qRT-PCR primers used to amplify the 16S RNA gene, cpe gene, and codY gene sequences were published previously (17,37). The primers used for abrB qRT-PCR were qabrBF and qabrBR (Table 2), and those used for qRT-PCR amplification of virX transcripts were qvirXF and qvirXR (Table 2).…”

Section: Methodsmentioning

confidence: 99%

“…In this regard, it is notable that typical type A food-poisoning strains, like SM101, are genotypically distinct from most other C. perfringens strains, as assessed by multilocus sequence type analyses of housekeeping genes (36). In addition, these typical food-poisoning strains carry a chromosomal cpe gene (rather than no cpe gene or a plasmid-borne cpe gene) but lack pfo and nanI genes, encoding perfringolysin O and NanI sialidase, respectively (30,37). They also produce an unusual small acidsoluble protein 4 (SASP4) variant that results in extremely tough spores highly resistant to heat, cold, and food preservatives compared to the spores of most other C. perfringens strains (38).…”

Section: Figmentioning

confidence: 99%

See 1 more Smart Citation

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

Freedman

Evans

et al. 2017

Infect Immun

Self Cite

View full text Add to dashboard Cite

Clostridium perfringens type D strains cause enterotoxemia and enteritis in livestock via epsilon toxin production. In type D strain CN3718, CodY was previously shown to increase the level of epsilon toxin production and repress sporulation. C. perfringens type A strains producing C. perfringens enterotoxin (CPE) cause human food poisoning and antibiotic-associated diarrhea. Sporulation is critical for C. perfringens type A food poisoning since spores contribute to transmission and resistance in the harsh food environment and sporulation is essential for CPE production. Therefore, the current study asked whether CodY also regulates sporulation and CPE production in SM101, a derivative of C. perfringens type A food-poisoning strain NCTC8798. An isogenic codY-null mutant of SM101 showed decreased levels of spore formation, along with lower levels of CPE production. A complemented strain recovered wild-type levels of both sporulation and CPE production. When this result was coupled with the earlier results obtained with CN3718, it became apparent that CodY regulation of sporulation varies among different C. perfringens strains. Results from quantitative reverse transcriptase PCR analysis clearly demonstrated that, during sporulation, codY transcript levels remained high in SM101 but rapidly declined in CN3718. In addition, abrB gene expression patterns varied significantly between codY-null mutants of SM101 and CN3718. Compared to the levels in their wild-type parents, the level of abrB gene expression decreased in the CN3718 codYnull mutant strain but significantly increased in the SM101 codY-null mutant strain, demonstrating CodY-dependent regulation differences in abrB expression between these two strains. This difference appears to be important since overexpression of the abrB gene in SM101 reduced the levels of sporulation and enterotoxin production, supporting the involvement of AbrB repression in regulating C. perfringens sporulation.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

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

Freedman

Evans

et al. 2017

Infect Immun

Self Cite

View full text Add to dashboard Cite

show abstract

“…Compared to SB, NADNA is an even more effective inhibitor of sialidase activity in culture supernatants, yet NADNA has little effect on the sialidase activity in C. perfringens cultures. Consequently, NADNA has no effect on ETX production and also causes weaker inhibition of Caco-2 cell adherence (34). Why NADNA has limited function in vivo requires future investigation.…”

Section: Discussionmentioning

confidence: 99%

“…The two larger sialidases, NanJ (129 kDa) and NanI (77 kDa), are secreted from C. perfringens, while the smallest enzyme, NanH (43 kDa), is located in the cytoplasm of log-phase cultures but can also appear extracellularly in older cultures (32,33). A recent study (32) found that type D strain CN3718 produces all three sialidases, with NanI being the major secreted sialidase of this strain, as it is for many other C. perfringens strains (34,35). NanI has an emerging potential role in pathogenesis.…”

mentioning

confidence: 99%

NanI Sialidase, CcpA, and CodY Work Together To Regulate Epsilon Toxin Production by Clostridium perfringens Type D Strain CN3718

Freedman

McClane

2015

J Bacteriol

Self Cite

View full text Add to dashboard Cite

Clostridium perfringens type D strains are usually associated with diseases of livestock, and their virulence requires the production of epsilon toxin (ETX). We previously showed (J. Li, S. Sayeed, S. Robertson, J. Chen, and B. A. McClane, PLoS Pathog 7:e1002429, 2011, http://dx.doi.org/10.1371/journal.ppat.1002429) that BMC202, a nanI null mutant of type D strain CN3718, produces less ETX than wild-type CN3718 does. The current study proved that the lower ETX production by strain BMC202 is due to nanI gene disruption, since both genetic and physical (NanI or sialic acid) complementation increased ETX production by BMC202. Furthermore, a sialidase inhibitor that interfered with NanI activity also reduced ETX production by wild-type CN3718. The NanI effect on ETX production was shown to involve reductions in codY and ccpA gene transcription levels in BMC202 versus wild-type CN3718. Similar to CodY, CcpA was found to positively control ETX production. A double codY ccpA null mutant produced even less ETX than a codY or ccpA single null mutant. CcpA bound directly to sequences upstream of the etx or codY start codon, and bioinformatics identified putative CcpA-binding cre sites immediately upstream of both the codY and etx start codons, suggesting possible direct CcpA regulatory effects. A ccpA mutation also decreased codY transcription, suggesting that CcpA effects on ETX production can be both direct and indirect, including effects on codY transcription. Collectively, these results suggest that NanI, CcpA, and CodY work together to regulate ETX production, with NanI-generated sialic acid from the intestines possibly signaling type D strains to upregulate their ETX production and induce disease. IMPORTANCEClostridium perfringens NanI was previously shown to increase ETX binding to, and cytotoxicity for, MDCK host cells. The current study demonstrates that NanI also regulates ETX production via increased transcription of genes encoding the CodY and CcpA global regulators. Results obtained using single ccpA or codY null mutants and a ccpA codY double null mutant showed that codY and ccpA regulate ETX production independently of one another but that ccpA also affects codY transcription. Electrophoretic mobility shift assays and bioinformatic analyses suggest that both CodY and CcpA may directly regulate etx transcription. Collectively, results of this study suggest that sialic acid generated by NanI from intestinal sources signals ETX-producing C. perfringens strains, via CcpA and CodY, to upregulate ETX production and cause disease. C lostridium perfringens is an important cause of human and livestock infections, including many diseases originating in the intestines (1, 2). The virulence of C. perfringens is largely attributable to its prolific toxin-producing ability, with different toxins involved in specific diseases (1,3,4). By definition, type D strains must produce both C. perfringens alpha toxin (CPA) and epsilon toxin (ETX). ETX is a class B NIAID priority toxin, since it is one of the most potent ba...

show abstract

“…Wild-type C. perfringens type A to E isolates used in this study are listed and described in Table 1. The toxin genotypes (A to E) and some phenotypic characteristics of these isolates were determined previously (23,24,(26)(27)(28)(29)(30)(31)(32)(33)(34). All C. perfringens isolates used in this study were maintained as stock cultures in cooked meat medium (Oxoid) and stored at Ϫ20°C.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Clostridium perfringens TpeL Toxin Gene Carriage, Production, Cytotoxic Contributions, and Trypsin Sensitivity

Chen

McClane

2015

Infect Immun

Self Cite

View full text Add to dashboard Cite

Large clostridial toxins (LCTs) are produced by at least four pathogenic clostridial species, and several LCTs are proven pivotal virulence factors for both human and veterinary diseases. TpeL is a recently identified LCT produced by Clostridium perfringens that has received relatively limited study. In response, the current study surveyed carriage of the tpeL gene among different C. perfringens strains, detecting this toxin gene in some type A, B, and C strains but not in any type D or E strains. This study also determined that all tested strains maximally produce, and extracellularly release, TpeL at the late-log or early-stationary growth stage during in vitro culture, which is different from the maximal late-stationary-phase production reported previously for other LCTs and for TpeL production by C. perfringens strain JIR12688. In addition, the present study found that TpeL levels in culture supernatants can be repressed by either glucose or sucrose. It was also shown that, at natural production levels, TpeL is a significant contributor to the cytotoxic activity of supernatants from cultures of tpeL-positive strain CN3685. Lastly, this study identified TpeL, which presumably is produced in the intestines during diseases caused by TpeL-positive type B and C strains, as a toxin whose cytotoxicity decreases after treatment with trypsin; this finding may have pathophysiologic relevance by suggesting that, like beta toxin, TpeL contributes to type B and C infections in hosts with decreased trypsin levels due to disease, diet, or age.

show abstract

Contributions of NanI Sialidase to Caco-2 Cell Adherence by Clostridium perfringens Type A and C Strains Causing Human Intestinal Disease

Cited by 41 publications

References 43 publications

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

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

NanI Sialidase, CcpA, and CodY Work Together To Regulate Epsilon Toxin Production by Clostridium perfringens Type D Strain CN3718

Characterization of Clostridium perfringens TpeL Toxin Gene Carriage, Production, Cytotoxic Contributions, and Trypsin Sensitivity

Contact Info

Product

Resources

About