Pili of <i>Vibrio cholerae</i> O1 Biotype El Tor: A Comparative Study on Adhesive and Non‐Adhesive Strains

Iwanaga, Masaaki; Nakasone, Noboru; Ehara, Masahiko

doi:10.1111/j.1348-0421.1989.tb01492.x

Cited by 20 publications

(20 citation statements)

References 17 publications

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“…11 and 12; Fig. 1 30°C and AKI represents growth under conditions previously described (11).] Whole cell lysates were prepared by boiling cell pellets in SDS/PAGE sample buffer.…”

Section: Resultsmentioning

confidence: 99%

“…Growth of V cholerae in LB was done overnight at 30°C on a tube roller. AKI conditions were as described (11,13). Ampicillin and kanamycin were used at final concentrations of 100 ,ug/ml and 30 ,ug/ml, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Classical strains express high levels of toxin and TCP in standard laboratory medium when cultivated at 25-30°C (5, 9, 10). El Tor strains, by contrast, require growth under special conditions, termed AKI, for high-level expression of ToxRregulated genes (11,12). Under the AKI conditions, organisms are cultivated for several hours in bicarbonate without aeration, followed by several hours of growth with vigorous aeration (11,12,28).…”

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confidence: 99%

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Differential expression of the ToxR regulon in classical and E1 Tor biotypes of Vibrio cholerae is due to biotype-specific control over toxT expression.

DiRita

Neely

Taylor

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

106

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The two major disease-causing biotypes of Vibrio cholerae, classical and El Tor, exhibit differences in their epidemic nature. Their behavior in the laboratory also differs in that El Tor strains produce two major virulence factors, cholera toxin (CT) and the toxin coregulated pilus (TCP), only under very restricted growth conditions, whereas classical strains do so in standard laboratory medium. Expression of toxin and TCP is controlled by two activator proteins, ToxR and ToxT, that operate in cascade fashion with ToxR controlling the synthesis of ToxT. Both biotypes express equivalent levels of ToxR, but only classical strains appear to express ToxT when grown in standard medium. In this report we show that restrictive expression of CT and TCP can be overcome in El Tor strains by expressing toxT independently of ToxR. An El Tor strain lacking functional ToxT does not express CT or TCP, ruling out existence of a cryptic pathway for virulence regulation in this biotype. These results may have implications for understanding the evolution of El Tor strains toward reduced virulence with respect to classical strains.

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“…11 and 12; Fig. 1 30°C and AKI represents growth under conditions previously described (11).] Whole cell lysates were prepared by boiling cell pellets in SDS/PAGE sample buffer.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Differential expression of the ToxR regulon in classical and E1 Tor biotypes of Vibrio cholerae is due to biotype-specific control over toxT expression.

DiRita

Neely

Taylor

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

106

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“…The molecular weight of the pilin (subunit protein of the pilus) from various organisms was about 20 kDa (11,13,14,22,33), and that of Ae6-R pili was 18 kDa. When the Ae6-R pili sample without the addition of 2ME was applied to SDS-PAGE, the migration was delayed and protein band appeared at 36 kDa.…”

Section: Discussionmentioning

confidence: 99%

Pili of Aeromonas hydrophila: Purification, Characterization, and Biological Role

Honma

Nakasone

1990

Microbiology and Immunology

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Aeromonas hydrophila (Ae6) has 2 morphologically distinctive kinds of pili. One appeared rigid, channeled, and straight with a diameter of 9 nm (Ae6-R pili). The other looked flexible, wavy, and having helical structure with a diameter of 7 nm (Ae6-W pili). Ae6-R pili were purified and characterized.The pili consisted of a subunit protein with a molecular weight of 18 kDa as estimated by SDS-PAGE, and contained 42.3% hydrophobic amino acids and one cysteine residue.The pilus was solubilized to 18 kDa subunit protein by 2-mercaptoethanol.dithiothreitol, hydrochloric acid, or heating at 120 C for 5 min. The organism Ae6 was strongly adhesive to rabbit intestines as well as human intestines, and agglutinated erythrocytes. Anti-pili antibody (Fab fraction) did not block the adhesion. Purified Ae6-R pili did not adhere to the intestine or to the erythrocytes.However, the anti-pili Fab inhibited pellicle formation of the organisms cultured in broth, and also inhibited salt agglutination with ammonium sulfate.From these results, Ae6-R pili are not likely a colonization factor but probably play a role in the autoaggregation of the organisms.

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“…cholerae without regard to serogroup, and have isolated several flexible pili (14,27,42,43). Among them, the one isolated from V. cholerae 034, strain NAGVI4, adhered to rabbit intestinal epithelium.…”

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Gene Analysis of Vibrio cholerae NAGV14 Pilus and Its Distribution

Kuroki

Toma

Nakasone

et al. 2001

Microbiology and Immunology

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Adhesive pilus of Vibrio cholerae 034, strain NAGVI4, was genetically analyzed. The deduced amino acid (aa) sequence of the major pilin structural gene (VcfA) was 67% homologous to the MshA pilin in the N-terminal region, but no homology was found in the C-terminal region which contained the antigenic epitopes. Upstream and downstream flanking regions examined were highly homologous to mshB and mshC of the MSHA (mannose-sensitive hemagglutinin) gene locus. A short leader sequence and a pair of cysteines near the C-terminus which are the characteristics of type 4a pilus family were found. The major pilin structural gene of NAGV14 was compared to that of a strain VI0 producing non-adhesive pili. The deduced aa sequences showed 60% homology, and the distance between two cysteines in the C-terminal region was different. A total of 177 V. cholerae strains were investigated for the presence of a type 4 pilus gene locus by PCR, and 95% were positive. The major pilin gene of NAGV14 was detected in 4 of 93 V. cholerae non-Ol, non-0139 strains tested, but none of the V. cholerae 01 and 0139 (72 and 12 strains, respectively). Our result suggested that a type 4 pilus gene locus similar to the MSHA gene locus is widely distributed among V. cholerae strains. We proposed naming this type 4 pilus gene locus the VCF (for V. cholerae flexible pili) gene locus.

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Pili of Vibrio cholerae O1 Biotype El Tor: A Comparative Study on Adhesive and Non‐Adhesive Strains

Cited by 20 publications

References 17 publications

Differential expression of the ToxR regulon in classical and E1 Tor biotypes of Vibrio cholerae is due to biotype-specific control over toxT expression.

Differential expression of the ToxR regulon in classical and E1 Tor biotypes of Vibrio cholerae is due to biotype-specific control over toxT expression.

Pili of Aeromonas hydrophila: Purification, Characterization, and Biological Role

Gene Analysis of Vibrio cholerae NAGV14 Pilus and Its Distribution

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