Identification of new loci involved in adhesion of to eukaryotic cells

The GenBank accession numbers for the sequences determined in this work are AF104224–AF104229.

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Abstract

Insertional mutagenesis was performed with Tn in the genetic background of an deletion mutant to identify new adhesion determinants in . Four insertion mutants defective in adhesion to eukaryotic cells were identified. Insertion sites were cloned by inverse-PCR and sequenced. The genetic organization of insertion regions was further analysed by screening and sequencing DNA fragments from a dIII library and by searching databases. Three adhesion-defective mutants each had one copy of Tn inserted into their chromosome. The insertion sites were different in the three mutants: (i) upstream from two ORFs in tandem, similar to and of , respectively; (ii) within an ORF encoding a putative 126 amino-acid-polypeptide with no significant similarity to any known protein; (iii) within an ORF similar to a ORF with no known function, just upstream from an operon similar to an ABC (ATP-binding cassette) transporter operon from . The excisants obtained from these mutants using the excision reporter plasmid pTCR9 recovered full adhesion capacity. A fourth mutant was the most severely defective in adhesion. It had five Tn insertions, one of which was upstream from and , and another of which was upstream from , a gene encoding a surface-exposed autolysin with a C terminus similar to that of InlB. Ami was clearly involved because an null mutant constructed in an EGDΔ background was adhesion-defective. Thus new regions involved in the adhesion of to eukaryotic cells were identified. Further study is required to define more accurately the roles of these regions in the adhesion process itself.

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2000-03-01
2024-03-28
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