Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111

Birgit Huber; Kathrin Riedel; Manuela Köthe; Michael Givskov; Søren Molin; Leo Eberl

Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111

Birgit Huber, Kathrin Riedel, Manuela Köthe, Michael Givskov, Søren Molin, Leo Eberl

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

133 Citationer (Scopus)

Abstract

Udgivelsesdato: 2002-Oct

Originalsprog	Engelsk
Tidsskrift	Molecular Microbiology
Vol/bind	46
Udgave nummer	2
Sider (fra-til)	411-26
Antal sider	15
ISSN	0950-382X
Status	Udgivet - 2002
Udgivet eksternt	Ja

Bibliografisk note

Keywords: 4-Butyrolactone; Bacterial Proteins; Biofilms; Burkholderia Infections; Burkholderia cepacia; Cystic Fibrosis; DNA Transposable Elements; Gene Expression Regulation, Bacterial; Humans; Hydrophobicity; Image Processing, Computer-Assisted; Microscopy, Confocal; Mutagenesis, Insertional; Polystyrenes; Signal Transduction; Surface Properties

Citationsformater

@article{a4828d00fcf111ddb219000ea68e967b,

title = "Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111",

abstract = "Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, we report the isolation of 13 random mini-Tn5 insertion mutants of B. cepacia H111 that are defective in biofilm formation on a polystyrene surface. We show that the screening procedure used in this study is biased towards mutants defective in the late stages of biofilm development. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains revealed that the isolated mutants are impaired in their abilities to develop a typical three-dimensional biofilm structure. Molecular investigations showed that the genes required for biofilm maturation fall into several classes: (i). genes encoding for surface proteins; (ii). genes involved in the biogenesis and maintenance of an integral outer membrane; and (iii). genes encoding regulatory factors. It is shown that three of the regulatory mutants produce greatly reduced amounts of N-octanoylhomoserine lactone (C8-HSL). This compound serves as the major signal molecule of the cep quorum-sensing system. As this density-dependent regulatory system is involved in the regulation of biofilm maturation, we investigated the interplay between the three regulatory genes and the quorum-sensing cascade. The results of these investigations show that the identified genes encode for regulatory elements that are positioned upstream of the cep system, indicating that the quorum-sensing system of B. cepacia is a major checkpoint for biofilm formation.",

author = "Birgit Huber and Kathrin Riedel and Manuela K{\"o}the and Michael Givskov and S{\o}ren Molin and Leo Eberl",

note = "Keywords: 4-Butyrolactone; Bacterial Proteins; Biofilms; Burkholderia Infections; Burkholderia cepacia; Cystic Fibrosis; DNA Transposable Elements; Gene Expression Regulation, Bacterial; Humans; Hydrophobicity; Image Processing, Computer-Assisted; Microscopy, Confocal; Mutagenesis, Insertional; Polystyrenes; Signal Transduction; Surface Properties",

year = "2002",

language = "English",

volume = "46",

pages = "411--26",

journal = "Molecular Microbiology",

issn = "0950-382X",

publisher = "Wiley-Blackwell",

number = "2",

}

TY - JOUR

T1 - Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111

AU - Huber, Birgit

AU - Riedel, Kathrin

AU - Köthe, Manuela

AU - Givskov, Michael

AU - Molin, Søren

AU - Eberl, Leo

N1 - Keywords: 4-Butyrolactone; Bacterial Proteins; Biofilms; Burkholderia Infections; Burkholderia cepacia; Cystic Fibrosis; DNA Transposable Elements; Gene Expression Regulation, Bacterial; Humans; Hydrophobicity; Image Processing, Computer-Assisted; Microscopy, Confocal; Mutagenesis, Insertional; Polystyrenes; Signal Transduction; Surface Properties

PY - 2002

Y1 - 2002

N2 - Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, we report the isolation of 13 random mini-Tn5 insertion mutants of B. cepacia H111 that are defective in biofilm formation on a polystyrene surface. We show that the screening procedure used in this study is biased towards mutants defective in the late stages of biofilm development. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains revealed that the isolated mutants are impaired in their abilities to develop a typical three-dimensional biofilm structure. Molecular investigations showed that the genes required for biofilm maturation fall into several classes: (i). genes encoding for surface proteins; (ii). genes involved in the biogenesis and maintenance of an integral outer membrane; and (iii). genes encoding regulatory factors. It is shown that three of the regulatory mutants produce greatly reduced amounts of N-octanoylhomoserine lactone (C8-HSL). This compound serves as the major signal molecule of the cep quorum-sensing system. As this density-dependent regulatory system is involved in the regulation of biofilm maturation, we investigated the interplay between the three regulatory genes and the quorum-sensing cascade. The results of these investigations show that the identified genes encode for regulatory elements that are positioned upstream of the cep system, indicating that the quorum-sensing system of B. cepacia is a major checkpoint for biofilm formation.

AB - Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, we report the isolation of 13 random mini-Tn5 insertion mutants of B. cepacia H111 that are defective in biofilm formation on a polystyrene surface. We show that the screening procedure used in this study is biased towards mutants defective in the late stages of biofilm development. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains revealed that the isolated mutants are impaired in their abilities to develop a typical three-dimensional biofilm structure. Molecular investigations showed that the genes required for biofilm maturation fall into several classes: (i). genes encoding for surface proteins; (ii). genes involved in the biogenesis and maintenance of an integral outer membrane; and (iii). genes encoding regulatory factors. It is shown that three of the regulatory mutants produce greatly reduced amounts of N-octanoylhomoserine lactone (C8-HSL). This compound serves as the major signal molecule of the cep quorum-sensing system. As this density-dependent regulatory system is involved in the regulation of biofilm maturation, we investigated the interplay between the three regulatory genes and the quorum-sensing cascade. The results of these investigations show that the identified genes encode for regulatory elements that are positioned upstream of the cep system, indicating that the quorum-sensing system of B. cepacia is a major checkpoint for biofilm formation.

M3 - Journal article

C2 - 12406218

SN - 0950-382X

VL - 46

SP - 411

EP - 426

JO - Molecular Microbiology

JF - Molecular Microbiology

IS - 2

ER -