Identification of bacterial factors involved in type 1 fimbria expression using an Escherichia coli K12 proteome chip

Yi Wen Chen, Ching-Hao Teng, Yu Hsuan Ho, Tien Yu Jessica Ho, Wen Chun Huang, Masayuki Hashimoto, I. Yuan Chiang, Chien-Sheng Chen

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Type 1 fimbriae are filamentous structures on Escherichia coli. These structures are important adherence factors. Because binding to the host cells is the first step of infection, type 1 fimbria is an important virulence factor of pathogenic E. coli. Expression of type 1 fimbria is regulated by a phase variation in which each individual bacterium can alternate between fimbriated (phase-ON) and nonfimbriated (phase-OFF) states. The phase variation is regulated by the flipping of the 314-bp fimS fragment, which contains the promoter driving the expression of the genes required for the synthesis of type 1 fimbria. Thus, the bacterial proteins able to interact with fimS are likely to be involved in regulating the expression of type 1 fimbria. To identify novel type 1 fimbria-regulating factors, we used an E. coli K12 proteome chip to screen for the bacterial factors able to interact with a 602-bp DNA fragment containing fimS and its adjacent regions. The Spr protein was identified by the proteome chip-based screening and further confirmed to be able to interact with fimS by electrophoretic mobility shift assay. Deletion of spr in the neonatal meningitis E. coli strain RS218 significantly increased the ratio of the bacterial colonies that contained the type 1 fimbria phase-ON cells on agar plates. In addition, Spr interfered with the interactions of fimS with the site-specific recombinases, FimB and FimE, which are responsible for mediating the flipping of fimS. These results suggest that Spr is involved in the regulation of type 1 fimbria expression through direct interaction with the invertible element fimS. These findings facilitate our understanding of the regulation of type 1 fimbria.

Original languageEnglish
Pages (from-to)1485-1494
Number of pages10
JournalMolecular and Cellular Proteomics
Volume13
Issue number6
DOIs
Publication statusPublished - 2014 Jan 1

Fingerprint

Escherichia coli K12
Proteome
Escherichia coli
Escherichia coli Meningitis
Bacterial Proteins
Electrophoretic Mobility Shift Assay
Virulence Factors
Agar
Bacteria
Gene Expression
Electrophoretic mobility
DNA
Infection
Assays
Screening
Proteins
Genes
Site-specific recombinase

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

Cite this

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title = "Identification of bacterial factors involved in type 1 fimbria expression using an Escherichia coli K12 proteome chip",
abstract = "Type 1 fimbriae are filamentous structures on Escherichia coli. These structures are important adherence factors. Because binding to the host cells is the first step of infection, type 1 fimbria is an important virulence factor of pathogenic E. coli. Expression of type 1 fimbria is regulated by a phase variation in which each individual bacterium can alternate between fimbriated (phase-ON) and nonfimbriated (phase-OFF) states. The phase variation is regulated by the flipping of the 314-bp fimS fragment, which contains the promoter driving the expression of the genes required for the synthesis of type 1 fimbria. Thus, the bacterial proteins able to interact with fimS are likely to be involved in regulating the expression of type 1 fimbria. To identify novel type 1 fimbria-regulating factors, we used an E. coli K12 proteome chip to screen for the bacterial factors able to interact with a 602-bp DNA fragment containing fimS and its adjacent regions. The Spr protein was identified by the proteome chip-based screening and further confirmed to be able to interact with fimS by electrophoretic mobility shift assay. Deletion of spr in the neonatal meningitis E. coli strain RS218 significantly increased the ratio of the bacterial colonies that contained the type 1 fimbria phase-ON cells on agar plates. In addition, Spr interfered with the interactions of fimS with the site-specific recombinases, FimB and FimE, which are responsible for mediating the flipping of fimS. These results suggest that Spr is involved in the regulation of type 1 fimbria expression through direct interaction with the invertible element fimS. These findings facilitate our understanding of the regulation of type 1 fimbria.",
author = "Chen, {Yi Wen} and Ching-Hao Teng and Ho, {Yu Hsuan} and Ho, {Tien Yu Jessica} and Huang, {Wen Chun} and Masayuki Hashimoto and Chiang, {I. Yuan} and Chien-Sheng Chen",
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Identification of bacterial factors involved in type 1 fimbria expression using an Escherichia coli K12 proteome chip. / Chen, Yi Wen; Teng, Ching-Hao; Ho, Yu Hsuan; Ho, Tien Yu Jessica; Huang, Wen Chun; Hashimoto, Masayuki; Chiang, I. Yuan; Chen, Chien-Sheng.

In: Molecular and Cellular Proteomics, Vol. 13, No. 6, 01.01.2014, p. 1485-1494.

Research output: Contribution to journalArticle

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AU - Teng, Ching-Hao

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AB - Type 1 fimbriae are filamentous structures on Escherichia coli. These structures are important adherence factors. Because binding to the host cells is the first step of infection, type 1 fimbria is an important virulence factor of pathogenic E. coli. Expression of type 1 fimbria is regulated by a phase variation in which each individual bacterium can alternate between fimbriated (phase-ON) and nonfimbriated (phase-OFF) states. The phase variation is regulated by the flipping of the 314-bp fimS fragment, which contains the promoter driving the expression of the genes required for the synthesis of type 1 fimbria. Thus, the bacterial proteins able to interact with fimS are likely to be involved in regulating the expression of type 1 fimbria. To identify novel type 1 fimbria-regulating factors, we used an E. coli K12 proteome chip to screen for the bacterial factors able to interact with a 602-bp DNA fragment containing fimS and its adjacent regions. The Spr protein was identified by the proteome chip-based screening and further confirmed to be able to interact with fimS by electrophoretic mobility shift assay. Deletion of spr in the neonatal meningitis E. coli strain RS218 significantly increased the ratio of the bacterial colonies that contained the type 1 fimbria phase-ON cells on agar plates. In addition, Spr interfered with the interactions of fimS with the site-specific recombinases, FimB and FimE, which are responsible for mediating the flipping of fimS. These results suggest that Spr is involved in the regulation of type 1 fimbria expression through direct interaction with the invertible element fimS. These findings facilitate our understanding of the regulation of type 1 fimbria.

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