PilC of Neisseria meningitidis is involved in class II pilus formation and restores pilus assembly, natural transformation competence and adherence to epithelial cells in PilC-deficient gonococci.
Type 4 pili produced by the pathogenic Neisseria species constitute primary determinants for the adherence to host tissues. In addition to the major pilin subunit (PilE), neisserial pili contain the variable PilC proteins represented by two variant gene copies in most pathogenic Neisseria isolates. Based upon structural differences in the conserved regions of PilE, two pilus classes can be distinguished in Neisseria meningitidis. For class I pili found in both Neisseria gonorrhoeae and N. meningitidis, PilC proteins have been implicated in pilus assembly, natural transformation competence and adherence to epithelial cells. In this study, we used primers specific for the pilC2 gene of N.gonorrhoeae strain MS11 to amplify, by the polymerase chain reaction, and clone a homologous pilC gene from N. meningitidis strain A1493 which produces class II pili. This gene was sequenced and the deduced amino acid sequence showed 75.4% and 73.8% identity with the gonococcal PilC1 and PilC2, respectively. These values match the identity value of 74.1% calculated for the two N. gonorrhoeae MS11 PilC proteins, indicating a horizontal relationship between the N. gonorrhoeae and N. meningitidis pilC genes. We provide evidence that PilC functions in meningococcal class II pilus assembly and adherence. Furthermore, expression of the cloned N. meningitidis pilC gene in a gonococcal pilC1,2 mutant restores pilus assembly, adherence to ME-180 epithelial cells, and transformation competence to the wild-type level. Thus, PilC proteins exhibit indistinguishable functions in the context of class I and class II pili.
Ryll
R R
RR
Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Tübingen, Germany.
Rudel
T
T
Scheuerpflug
I
I
Barten
R
R
Meyer
T F
TF
D000595:Amino Acid Sequence; D001422:Bacterial Adhesion / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology*; D015139:Blotting, Southern; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis; D010450:Endopeptidases / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000235:genetics; D009345:Neisseria meningitidis / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes; D010957:Plasmids; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014169:Transformation, Bacterial / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009692:Nucleic Acid Heteroduplexes; C079208:pilC protein, Neisseria gonorrhoeae; D036781:Fimbriae Proteins; D010450:Endopeptidases