Isolation of a putative fimbrial adhesin from Bordetella pertussis and the identification of its gene.
We report the purification of a minor Bordetella pertussis fimbrial subunit, designated FimD, and the identification of its gene (fimD). FimD could be purified from the bulk of major fimbrial subunits by exploiting the fact that major subunit-subunit interactions are more stable in the presence of SDS than minor-major subunit interactions. To locate the gene for FimD, internal peptides of FimD were generated, purified and sequenced. Subsequently, an oligonucleotide probe, based on the primary sequence of one peptide, was used to clone fimD. The primary structure of FimD, derived from the DNA sequence of its gene, showed homology with a number of fimbrial adhesins. Most pronounced homology was observed with MrkD, a fimbrial adhesin derived from Klebsiella pneumoniae. These observations suggest that FimD may represent a B. pertussis fimbrial adhesin. With a fimD-specific probe we detected the presence of a fimD homologue in Bordetella parapertussis and Bordetella bronchiseptica but not in Bordetella avium. Cloning and sequencing revealed that the B. parapertussis and B. bronchiseptica fimD product differed from the B. pertussis fimD product in 20 and 1 amino acid residues, respectively. Since B. bronchiseptica is normally not a human pathogen, but causes respiratory disease in a wide range of non-human mammalian species, this may suggest that FimD recognizes a receptor that is well conserved in mammalian species. An in-frame deletion in fimD completely abolished FimD expression and also affected the expression of the major subunits Fim2 and Fim3 suggesting that, in contrast to other adhesins that are minor components of fimbriae, FimD is required for formation of the fimbrial structure.
Willems
R J
RJ
Molecular Microbiology Unit, National Institute of Health and Environmental Protection, Bilthoven, The Netherlands.
Geuijen
C
C
van der Heide
H G
HG
Matheson
M
M
Robinson
A
A
Versluis
L F
LF
Ebberink
R
R
Theelen
J
J
Mooi
F R
FR
D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001426:Bacterial Proteins; D001483:Base Sequence; D001884:Bordetella / Q000235:genetics / Q000472:pathogenicity; D016950:Bordetella bronchiseptica / Q000235:genetics / Q000472:pathogenicity; D001886:Bordetella pertussis / Q000235:genetics* / Q000472:pathogenicity; D003001:Cloning, Molecular; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000737:chemistry; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000235:genetics; D015183:Restriction Mapping; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014774:Virulence
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; C489284:fimD protein, Bordetella; C063629:fimD protein, bacteria; D036781:Fimbriae Proteins