Isolation and characterization of a monoclonal antibody directed against type 1 fimbriae organelles from Escherichia coli

Eisenstein, B I; Clements, Janice; Dodd, D C

doi:10.1128/iai.42.1.333-340.1983

Cited by 26 publications

(16 citation statements)

References 25 publications

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“…Numerous studies have suggested that the various gene clusters have been lost during evolution as a pathoadaptive response to the host. Notably, laboratory growth methods consistently used to demonstrate fimbrial production in strains of E. coli (19, 20) were not successful for either lab strains and clinical isolates of Shigella (17, 18). Our control media analyses, in which the combination of glucose and bile salts were absent, confirmed many of these previous findings on the phenotypic level.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, Shigella is thought to have lost the ability to produce “traditional” E. coli adherence factors as the bacteria adapted to an intracellular lifestyle (2) due to three main reasons. First, Shigella grown in standard laboratory media lack structures upon transmission electron microscopy (TEM) (17, 18), unlike some strains of E. coli in which adherence factors are thought to be constitutively expressed (19, 20). Second, examination of Shigella genomes deposited in GenBank reveals that almost all adherence gene clusters, such as type 1 fimbriae (10, 21) and curli (22), contain at least one annotated pseudogene that is crucial for either the adherence factor structure or the assembly process (17, 23, 24).…”

Section: Introductionmentioning

confidence: 99%

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Shigella flexneri adherence factor expression in in vivo-like conditions

Chanin

Nickerson

Llanos-Chea

et al. 2019

Preprint

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26The Shigella species are Gram-negative, facultative intracellular pathogens that invade the 27 colonic epithelium and cause significant diarrheal disease. Despite extensive research on the pathogen, 28 comprehensive understanding of how Shigella initiates contact with epithelial cells remains unknown. 29 Shigella maintains many of the same Escherichia coli adherence gene operons; however, at least one 30 critical gene component in each operon is currently annotated as a pseudogene in reference genomes. 31 These annotations, coupled with a lack of structures upon microscopic analysis following growth in 32 laboratory media, have led the field to hypothesize that Shigella is unable to produce fimbriae or other 33 "traditional" adherence factors. Nevertheless, our previous analyses have demonstrated that a 34 combination of bile salts and glucose induce both biofilm formation and adherence to colonic epithelial 35 cells. Through a two-part investigation, we first utilized various transcriptomic analyses to demonstrate 36 that S. flexneri strain 2457T adherence gene operons are transcribed. Subsequently, we performed 37 mutation, electron microscopy, biofilm, infection, and proteomic analyses to characterize three of the 38 structural genes. In combination, these studies demonstrate that despite the gene annotations, S. 39 flexneri 2457T uses adherence factors to initiate biofilm formation as well as epithelial cell contact. 40 Furthermore, host factors, namely glucose and bile salts in the small intestine, offer key environmental 41 stimuli required for proper adherence factor expression in S. flexneri. This research may have a 42 significant impact on vaccine development for Shigella and further highlights the importance of 43 utilizing in vivo-like conditions to study bacterial pathogenesis. 44 45 Importance 46 Bacterial pathogens have evolved to regulate virulence gene expression at critical points in the 47 colonization and infection processes to successfully cause disease. The Shigella species infect the 48 epithelial cells lining the colon to result in millions of cases of diarrhea and a significant global health 49 burden. As antibiotic resistance rates increase, understanding the mechanisms of infection are vital to 50 3 ensure successful vaccine development. Despite significant gains in our understanding of Shigella 51 infection, it remains unknown how the bacteria initiate contact with the colonic epithelium. Most 52 pathogens harbor multiple adherence factors to facilitate this process, but Shigella was thought to have 53 lost the ability to produce these factors. Interestingly, we have identified conditions that mimic some 54 features of gastrointestinal transit and enable Shigella to express adherence factors. This work 55 highlights aspects of genetic regulation for Shigella adherence factors and may have a significant 56 impact on future vaccine development. 57 58 59Shigella flexneri is a Gram-negative, facultative anaerobe that infects millions of people each 60 year by causing water...

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Shigella flexneri adherence factor expression in in vivo-like conditions

Chanin

Nickerson

Llanos-Chea

et al. 2019

Preprint

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“…Chemicals and buffers. All reagents were as previously described (2,6) Antibodies. For the enzyme-linked immunosorbent assay (ELISA) inhibition assay, we used rabbit serum specific for type 1 fimbriae as previously described (2).…”

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confidence: 99%

“…For the enzyme-linked immunosorbent assay (ELISA) inhibition assay, we used rabbit serum specific for type 1 fimbriae as previously described (2). For immunoprecipitations, we used this serum, a monoclonal antibody (MAb) previously described (6), and monospecific rabbit anti-maltose-binding protein serum, kindly provided by Phil Bassford, Jr. (10). Rabbit antimouse serum was kindly provided by Judy Teale (6).…”

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confidence: 99%

“…We recovered the precipitate by centrifuging the solution for 3 min at 4°C in a Beckman Microfuge model B (Fullerton, Calif.). After washing the precipitate twice with ice-cold acetone, we solubilized it in 0.1 ml of a 1% SDS-50 mM Tris-hydrochloride (pH 8)-i mM EDTA solution and performed the immunoprecipitation as previously described (2,6). The TCA-acetone treatment did not affect the efficiency of immunoprecipitation or cause any detectable disaggregation of organelles into subunits.…”

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Kinetic analysis of the synthesis and assembly of type 1 fimbriae of Escherichia coli

Dodd¹,

Eisenstein²

1984

J Bacteriol

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The adhesive organelles (type 1 fimbriae) of K-12 and other isolates of Escherichia coli are composed of identical 17,000-dalton subunits. We examined the assembly of these subunits into fimbrial organelles. After synthesis, the nascent subunits were first processed and then assembled into the organelles; the assembly step took almost 3 min in log-phase cultures at 37°C. Even during blockage of protein synthesis, the free subunits continued to assemble until the pool was depleted. This pool was small in comparison with the amount of total fimbrial protein already assembled into surface organelles and was not sufficient to regenerate new detectable organelles after the removal of preexistent ones by blending. Assembly appeared to slow when the metabolic rate of the bacterial cells slowed, since subunits took longer to appear in the organelles at lower than optimal temperatures or as a culture entered the stationary phase. The synthetic rate of subunits slowed sooner than that of total cellular proteins as a culture approached the stationary phase and ceased completely as the culture entered the stationary phase. The amount of fimbrial antigen expressed on the surface of the cells remained relatively constant during growth of a culture. Type 1 fimbriae (or pili) are proteinaceous appendages 7 nm in width projecting from the surface of many gram-negative bacteria (4). They consist of aggregates of helically arranged identical 17,000-dalton polypeptides with 3.

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Regulation and Expression of Bacterial Adhesins

Ofek¹,

Doyle

1994

Bacterial Adhesion to Cells and Tissues

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Isolation and characterization of a monoclonal antibody directed against type 1 fimbriae organelles from Escherichia coli

Cited by 26 publications

References 25 publications

Shigella flexneri adherence factor expression in in vivo-like conditions

Shigella flexneri adherence factor expression in in vivo-like conditions

Kinetic analysis of the synthesis and assembly of type 1 fimbriae of Escherichia coli

Regulation and Expression of Bacterial Adhesins

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