The Longus type IV pilus of enterotoxigenic Escherichia coli (ETEC) mediates bacterial self-aggregation and protection from antimicrobial agents

DNA amplification of lngA, the structural gene of longus type IV pilus produced by human enterotoxigenicEscherichia coli (ETEC) was achieved by the use of specific oligonucleotide primers designed from the nucleotide sequence oflngA. A 630-bp fragment representing the entirelngA gene was amplified in eight prototype strains previously characterized as longus positive. Five ETEC strains producing colonization factor antigen III (CFA III) (also a type IV pilus) were also positive by PCR, confirming the DNA homology between CFA III and longus. None of the non-ETEC and non-E. colienteropathogens studied showed the 0.63-kbp amplicon. The procedure thus detected only ETEC strains harboring type IV pili genes with or without other colonization factors. Except for five lngAPCR-positive, probe-positive strains, all lngA PCR-positive strains produced the pilin as demonstrated by immunoblotting. To test the amplification procedure in a clinical setting, a collection of 264 fresh clinical E. coli strains isolated from 88 Mexican children with diarrhea was screened by PCR. Among 82 ETEC isolates found, 30 (36.5%) were lngA PCR-positive. Twenty-seven percent of the children shed ETEC that possessed lngA. In parallel with DNA probes or PCR protocols to detect enterotoxin genes, the lngA PCR method may prove useful for detection of ETEC harboring type IV pilus genes in epidemiological studies.

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Genetic Diversity of the Gene Cluster Encoding Longus, a Type IV Pilus of Enterotoxigenic Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.00722-07 ◽

2007 ◽

Vol 189 (24) ◽

pp. 9145-9149 ◽

Cited By ~ 22

Author(s):

Oscar G. Gomez-Duarte ◽

Sujay Chattopadhyay ◽

Scott J. Weissman ◽

Jorge A. Giron ◽

James B. Kaper ◽

...

Keyword(s):

Escherichia Coli ◽

Gene Cluster ◽

Protein Level ◽

Enterotoxigenic Escherichia Coli ◽

Subunit Gene ◽

Type Iv Pilus ◽

Strong Positive Selection ◽

Type Iv ◽

Structural Subunit ◽

Selection For

ABSTRACT Enterotoxigenic Escherichia coli (ETEC) strains produce a type IV pilus named Longus. We identified a 16-gene cluster involved in the biosynthesis of Longus that has 57 to 95% identity at the protein level to CFA/III, another type IV pilus of ETEC. Alleles of the Longus structural subunit gene lngA demonstrate a diversity of 12 to 19% at the protein level with strong positive selection for point replacements and horizontal transfer.

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Gene Cluster for Assembly of Pilus Colonization Factor Antigen III of Enterotoxigenic Escherichia coli

Infection and Immunity ◽

10.1128/iai.69.9.5864-5873.2001 ◽

2001 ◽

Vol 69 (9) ◽

pp. 5864-5873 ◽

Cited By ~ 33

Author(s):

Tooru Taniguchi ◽

Yukihiro Akeda ◽

Ayako Haba ◽

Yoko Yasuda ◽

Koichiro Yamamoto ◽

...

Keyword(s):

Escherichia Coli ◽

Nucleotide Sequence ◽

Gene Cluster ◽

Enterotoxigenic Escherichia Coli ◽

Human Colon Carcinoma Cell ◽

E Coli ◽

Colonization Factor ◽

Type Iv ◽

K 12 ◽

Factor Antigen

ABSTRACT The assembly of pilus colonization factor antigen III (CFA/III) of enterotoxigenic Escherichia coli (ETEC) requires the processing of CFA/III major pilin (CofA) by a prepilin peptidase (CofP), similar to other type IV pilus formation systems. CofA is produced initially as a 26.5-kDa preform pilin (prepilin) and then processed to a 20.5-kDa mature pilin by CofP which is predicted to be localized in the inner membrane. In the present experiment, we determined the nucleotide sequence of the whole region for CFA/III formation and identified a cluster of 14 genes, includingcofA and cofP. Several proteins encoded bycof genes were similar to previously described proteins, such as the toxin-coregulated pili of Vibrio cholerae and the bundle-forming pili of enteropathogenic E. coli. The G+C content of the cof gene cluster was 37%, which was significantly lower than the average for the E. coli genome (50%). The introduction of a recombinant plasmid containing thecof gene cluster into the E. coli K-12 strain conferred CFA/III biogenesis and the ability of adhesion to the human colon carcinoma cell line Caco-2. This is the first report of a complete nucleotide sequence of the type IV pili found in human ETEC, and our results provide a useful model for studying the molecular mechanism of CFA/III biogenesis and the role of CFA/III in ETEC infection.

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Deciphering the Conformational Dynamics of Escherichia coli Type IV Pilus

Biophysical Journal ◽

10.1016/j.bpj.2020.11.1885 ◽

2021 ◽

Vol 120 (3) ◽

pp. 294a

Author(s):

Yasaman Karami ◽

Olivera Francetic ◽

Nadia Izadi Pruneyre ◽

Michael Nilges

Keyword(s):

Escherichia Coli ◽

Conformational Dynamics ◽

Type Iv Pilus ◽

Type Iv

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Antimicrobial Resistance Genes in Enterotoxigenic Escherichia coli O149:K91 Isolates Obtained over a 23-Year Period from Pigs

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.47.10.3214-3221.2003 ◽

2003 ◽

Vol 47 (10) ◽

pp. 3214-3221 ◽

Cited By ~ 161

Author(s):

Christine Maynard ◽

John M. Fairbrother ◽

Sadjia Bekal ◽

François Sanschagrin ◽

Roger C. Levesque ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Antimicrobial Agents ◽

Enterotoxigenic Escherichia Coli ◽

Genotypic Resistance ◽

Class 1 Integron ◽

Phenotypic Resistance ◽

Antimicrobial Resistance Genes

ABSTRACT A total of 112 Escherichia coli O149:K91 strains isolated from pigs with diarrhea in Quebec, Canada, between 1978 and 2000 were characterized for their genotypic antimicrobial resistance profiles. Tests for resistance to 10 antimicrobial agents were conducted. Resistance to tetracycline and sulfonamides was found to be the most frequent, but resistance to cefotaxime and ceftiofur was absent. An increase in the number of isolates resistant to at least three antimicrobials was observed over time. The distribution of 28 resistance genes covering six antimicrobial families (beta-lactams, aminoglycosides, phenicols, tetracycline, trimethoprim, and sulfonamides) was assessed by colony hybridization. Significant differences in the distributions of tetracycline [tet(A), tet(B), tet(C)], trimethoprim (dhfrI, dhfrV, dhfrXIII), and sulfonamide (sulI, sulII) resistance genes were observed during the study period (1978 to 2000). Sixty percent of the isolates possessed a class 1 integron, illustrating the importance of integrons in the epidemiology of antibiotic resistance in E. coli strains from pigs. Amplification of the integron's variable region resulted in four distinct fragments of 1, 1.3, 1.6, and 1.8 kb, with the 1.6- and 1.8-kb fragments appearing only during the last half of the study period. Examination of linkages among the different resistance genes showed a variety of positive and negative associations. Association analysis of isolates divided into two groups, those isolated between 1978 and 1989 and those isolated between 1990 and 2000, revealed the appearance of new positive resistance gene associations. Our genotypic resistance analyses of ETEC isolates from pigs indicate that many of the antibiotic resistance genes behind phenotypic resistance are not static but, rather, are in a state of flux driven by various selection forces such as the use of specific antimicrobials.

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Type IV Longus Pilus of Enterotoxigenic Escherichia coli: Occurrence and Association with Toxin Types and Colonization Factors among Strains Isolated in Argentina

Journal of Clinical Microbiology ◽

10.1128/jcm.40.2.694-697.2002 ◽

2002 ◽

Vol 40 (2) ◽

pp. 694-697 ◽

Cited By ~ 14

Author(s):

M. G. Pichel ◽

N. Binsztein ◽

F. Qadri ◽

J. A. Giron

Keyword(s):

Escherichia Coli ◽

Enterotoxigenic Escherichia Coli ◽

Type Iv ◽

Colonization Factors

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Outer Membrane Targeting, Ultrastructure, and Single Molecule Localization of the Enteropathogenic Escherichia coli Type IV Pilus Secretin BfpB

Journal of Bacteriology ◽

10.1128/jb.06330-11 ◽

2012 ◽

Vol 194 (7) ◽

pp. 1646-1658 ◽

Cited By ~ 23

Author(s):

J. A. Lieberman ◽

N. A. Frost ◽

M. Hoppert ◽

P. J. Fernandes ◽

S. L. Vogt ◽

...

Keyword(s):

Escherichia Coli ◽

Outer Membrane ◽

Single Molecule ◽

Membrane Targeting ◽

Enteropathogenic Escherichia Coli ◽

Type Iv Pilus ◽

Type Iv

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BfpU, a soluble protein essential for type IV pilus biogenesis in enteropathogenic Escherichia coli

Microbiology ◽

10.1099/00221287-148-8-2507 ◽

2002 ◽

Vol 148 (8) ◽

pp. 2507-2518 ◽

Cited By ~ 14

Author(s):

Wiebke Schreiber ◽

Kelly D. Stone ◽

Margaret A. Strong ◽

Louis J. DeTolla, Jr ◽

Michael Hoppert ◽

...

Keyword(s):

Escherichia Coli ◽

Soluble Protein ◽

Enteropathogenic Escherichia Coli ◽

Type Iv Pilus ◽

Type Iv ◽

Pilus Biogenesis

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Characterization of a Novel Type IV Pilus Locus Encoded on the Large Plasmid of Locus of Enterocyte Effacement-Negative Shiga-Toxigenic Escherichia coli Strains That Are Virulent for Humans

Infection and Immunity ◽

10.1128/iai.70.6.3094-3100.2002 ◽

2002 ◽

Vol 70 (6) ◽

pp. 3094-3100 ◽

Cited By ~ 40

Author(s):

Potjanee Srimanote ◽

Adrienne W. Paton ◽

James C. Paton

Keyword(s):

Escherichia Coli ◽

Gastrointestinal Disease ◽

Large Plasmid ◽

Type Iv Pilus ◽

Upstream Gene ◽

Type Iv ◽

Uremic Syndrome ◽

Enterocyte Effacement

ABSTRACT The majority of Shiga-toxigenic Escherichia coli (STEC) strains isolated from humans with gastrointestinal disease carry large (approximately 90-kb) plasmids. We have been analyzing the megaplasmid (designated pO113) from an O113:H21 STEC strain (98NK2). This strain lacks the locus for enterocyte effacement (LEE) and yet was responsible for an outbreak of hemolytic uremic syndrome. In the present study, we demonstrate that pO113 carries a novel type IV pilus biosynthesis locus (pil) related to those of the IncI plasmids R721, R64, and ColIb9. The pO113 pil locus consists of 11 closely linked genes (pilL through pilV) with an additional separately transcribed upstream gene (pilI). It directs the expression of long thin pili on the 98NK2 surface and the hemagglutination of guinea pig erythrocytes. We also demonstrate that pO113 can be transferred by conjugation. However, the type IV pilus encoded by pO113 does not appear to be involved in the adherence of 98NK2 to HEp-2 or Hct-8 cells in vitro. Homologues of the pO113 pil locus were present in several other LEE-negative STEC strains but not in LEE-positive STEC strains belonging to serogroup O26, O111, or O157.

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