Identification of gene products involved in biofilm production by Moraxella catarrhalis ETSU-9 in vitro

Melanie M. Pearson, Eric J. Hansen

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Moraxella catarrhalis ETSU-9 was subjected to random transposon insertion mutagenesis to identify genes encoding products involved in the ability of the organism to form biofilms in vitro. Screening of approximately 3,000 transposon insertion mutants in the crystal violet-based biofilm assay system yielded six mutants that exhibited greatly reduced abilities to form biofilms. Three of these mutants had transposon insertions in the uspA2H gene, which encodes a surface protein previously shown to be involved in the ability of M. catarrhalis to both attach to human cell lines in vitro and resist killing by normal human serum. Random insertion mutagenesis of the uspA2H gene, involving the introduction of a 15-nucleotide fragment encoding 5 amino acids, was used to attempt to identify the domain(s) necessary for biofilm formation. Most of these insertions adversely affected biofilm formation, whereas the abilities of these same mutants to attach to Chang conjunctival epithelial cells in vitro were usually not reduced. Gain-of-function experiments showed that introduction of the M. catarrhalis ETSU-9 uspA2H gene into Escherichia coli conferred biofilm formation ability on this recombinant strain. Two of the other three M. catarrhalis ETSU-9 transposon insertion mutants that had greatly reduced abilities to form biofilms were shown to have insertions in genes encoding products predicted to be directly or indirectly involved in cell wall metabolism.

Original languageEnglish (US)
Pages (from-to)4316-4325
Number of pages10
JournalInfection and Immunity
Volume75
Issue number9
DOIs
StatePublished - Sep 2007

Fingerprint

Moraxella (Branhamella) catarrhalis
Biofilms
Genes
Mutagenesis
Organism Forms
Gentian Violet
Insertional Mutagenesis
In Vitro Techniques
Cell Wall
Membrane Proteins
Nucleotides
Epithelial Cells
Escherichia coli
Amino Acids
Cell Line
Serum

ASJC Scopus subject areas

  • Immunology

Cite this

Identification of gene products involved in biofilm production by Moraxella catarrhalis ETSU-9 in vitro. / Pearson, Melanie M.; Hansen, Eric J.

In: Infection and Immunity, Vol. 75, No. 9, 09.2007, p. 4316-4325.

Research output: Contribution to journalArticle

Pearson, Melanie M. ; Hansen, Eric J. / Identification of gene products involved in biofilm production by Moraxella catarrhalis ETSU-9 in vitro. In: Infection and Immunity. 2007 ; Vol. 75, No. 9. pp. 4316-4325.
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