Amino acid residue Ala-62 in the FimH fimbrial adhesin is critical for the adhesiveness of meningitis-associated Escherichia coli to collagens

Mol Microbiol. 1999 Mar;31(6):1747-57. doi: 10.1046/j.1365-2958.1999.01311.x.

Abstract

Adhesion of meningitis-associated Escherichia coli O18acK1H7 to collagens was characterized. The E. coli strain IHE 3034 adhered to type IV and type I collagens but not to type III collagen immobilized on glass. Collagens lack terminal mannosyl units, yet the bacterial adhesion was completely abolished in the presence of alpha-methyl-D-mannoside. A cat cassette was introduced into the filmA gene of IHE 3034, and the resulting mutant strain IHE 3034-2 failed to adhere to collagens. In contrast, insertion of a Gm cassette into the sfaA gene of IHE 3034, encoding the S-fimbrillin, had no significant effect on the adhesiveness. The fim cluster from IHE 3034 was cloned and expressed in trans in the fimA::cat mutant strain IHE 3034-2. The complemented strain IHE 3034-2(pRPO-1) exhibited adhesiveness to type IV and type I collagens, confirming the function of the type 1 fimbria in the adhesion. We have previously shown that the type 1 fimbria from E. coli K-12 strain PC31 does not confer bacterial adhesiveness to collagens. The fimH genes from E. coli IHE 3034 as well as from PC31 were expressed in the fimH-null strain MS4. The FimH from IHE 3034 potentiated collagen adherence, whereas the FimH from PC31 was inactive. Sequence comparison of fimH from IHE 3034 and PC31 revealed five amino-acid differences in the predicted mature FimH proteins: at residues 27, 62, 70, 78 and 201. Each of these residues in the IHE 3034-FimH were individually substituted to the corresponding amino acid in the PC31-FimH. The substitution S62-->A completely abolished collagen adhesiveness. The reverse substitution A62-->S in the PC31-FimH as well as in the FimH from another E. coli strain induced collagen adhesiveness to the level seen with IHE 3034-FimH. Out of nine fimH genes analysed from isolates of E. coli, collagen adhesiveness as well as alanine at position 62 in FimH were found only in two O18acK1H7 isolates with the isoenzyme profile ET type 1. Our results demonstrate that the amino-acid residue Ala-62 in the FimH lectin is critical for the adhesion to collagens by a highly virulent clonal group of E. coli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / chemistry*
  • Adhesins, Escherichia coli*
  • Animals
  • Bacterial Adhesion*
  • Bacterial Proteins / genetics
  • Collagen / metabolism*
  • Dose-Response Relationship, Drug
  • Escherichia coli / chemistry*
  • Escherichia coli / pathogenicity*
  • Escherichia coli Proteins*
  • Fimbriae Proteins*
  • Humans
  • Meningitis / microbiology*
  • Mice
  • Models, Genetic
  • Mutagenesis, Insertional
  • Phenotype
  • Placenta / metabolism
  • Point Mutation
  • Serum Albumin, Bovine / metabolism

Substances

  • Adhesins, Bacterial
  • Adhesins, Escherichia coli
  • Bacterial Proteins
  • Escherichia coli Proteins
  • fimH protein, E coli
  • ymcE protein, E coli
  • Fimbriae Proteins
  • Serum Albumin, Bovine
  • Collagen