Genetic characterization of Bordetella pertussis filamentous haemagglutinin: a protein processed from an unusually large precursor

The nucleotide sequence of the structural gene for filamentous haemagglutinin (FHA), fhaB, a crucial adherence factor for Bordetella perfussis, has been determined. Its 10774 nucleotides are far more than necessary to encode the 220kD biologically active, mature polypeptide product, suggesting a role for co‐or post‐translational processing. Fusion proteins derived from various portions of the fhaB open reading frame (ORF) were used to generate polyclonal antisera. Western immunoblot analysis of purified FHA and Bordetella sp. whole cell extracts with these antisera indicated that the 220kD product is encoded by the 5 portion of the ORF and that the smaller polypeptide species are breakdown products of this polypeptide. These data, as well as N‐terminal amino acid sequencing of the major polypeptide species, suggest a scheme for the proteolytic processing of an FHA precursor polypeptide.

[1]  D. Relman,et al.  Bordetella pertussis filamentous hemagglutinin: evaluation as a protective antigen and colonization factor in a mouse respiratory infection model , 1990, Infection and immunity.

[2]  S. Falkow,et al.  Analysis of Bordetella pertussis virulence gene regulation by use of transcriptional fusions in Escherichia coli , 1989, Journal of bacteriology.

[3]  R. Rappuoli,et al.  Sequences required for expression of Bordetella pertussis virulence factors share homology with prokaryotic signal transduction proteins. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[4]  V. Fischetti,et al.  Streptococcal M protein: molecular design and biological behavior , 1989, Clinical Microbiology Reviews.

[5]  R. Gibbons.  Bacterial Adhesion to Oral Tissues: A Model for Infectious Diseases , 1989, Journal of dental research.

[6]  C. Sasakawa,et al.  Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries , 1989, Molecular microbiology.

[7]  I. Charles,et al.  Molecular cloning and characterization of protective outer membrane protein P.69 from Bordetella pertussis. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Rappuoli,et al.  Filamentous hemagglutinin of Bordetella pertussis: nucleotide sequence and crucial role in adherence. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M. Lindberg,et al.  Nucleotide sequence of the gene for a fibronectin-binding protein from Staphylococcus aureus: use of this peptide sequence in the synthesis of biologically active peptides. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Danchin,et al.  Secretion of cyclolysin, the calmodulin‐sensitive adenylate cyclase‐haemolysin bifunctional protein of Bordetella pertussis. , 1988, The EMBO journal.

[11]  S. Redd,et al.  Immunoblot analysis of humoral immune responses following infection with Bordetella pertussis or immunization with diphtheria-tetanus-pertussis vaccine , 1988, Journal of clinical microbiology.

[12]  S. Falkow,et al.  Genetic analysis of a region of the Bordetella pertussis chromosome encoding filamentous hemagglutinin and the pleiotropic regulatory locus vir , 1988, Journal of bacteriology.

[13]  J. Horton,et al.  Identification of P1 gene domain containing epitope(s) mediating Mycoplasma pneumoniae cytoadherence , 1988, The Journal of experimental medicine.

[14]  A. Danchin,et al.  The calmodulin‐sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia col , 1988, Molecular microbiology.

[15]  C. Su,et al.  Cloning and sequence analysis of cytadhesin P1 gene from Mycoplasma pneumoniae , 1987, Infection and immunity.

[16]  E Ruoslahti,et al.  New perspectives in cell adhesion: RGD and integrins. , 1987, Science.

[17]  C. J. Duggleby,et al.  Cloning and nucleotide sequence analysis of the serotype 2 fimbrial subunit gene of Bordetella pertussis , 1987, Molecular microbiology.

[18]  C. Higgins,et al.  Sequence‐imposed structural constraints in the TonB protein of E. coli , 1986, FEBS letters.

[19]  M. Yeadon,et al.  1H-NMR study of mobility and conformational constraints within the proline-rich N-terminal of the LC1 alkali light chain of skeletal myosin. Correlation with similar segments in other protein systems. , 1986, European journal of biochemistry.

[20]  J. Vandekerckhove,et al.  Protein‐chemical analysis of pertussis toxin reveals homology between the subunits S2 and S3, between S1 and the A chains of enterotoxins of Vibrio cholerae and Escherichia coli and identifies S2 as the haptoglobin‐binding subunit , 1986, FEBS letters.

[21]  R. Rappuoli,et al.  Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Urisu,et al.  Filamentous hemagglutinin has a major role in mediating adherence of Bordetella pertussis to human WiDr cells , 1986, Infection and immunity.

[23]  H. Schaller,et al.  Characterization of foot-and-mouth disease virus gene products with antisera against bacterially synthesized fusion proteins , 1986, Journal of virology.

[24]  V. Fischetti,et al.  Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. , 1986, The Journal of biological chemistry.

[25]  E. Hewlett,et al.  Virulence factors of Bordetella pertussis. , 1986, Annual review of microbiology.

[26]  S. Falkow,et al.  The construction of a cloning vector designed for gene replacement in Bordetella pertussis. , 1986, Gene.

[27]  E. Tuomanen,et al.  Characterization of two adhesins of Bordetella pertussis for human ciliated respiratory-epithelial cells. , 1985, The Journal of infectious diseases.

[28]  C. Manclark,et al.  Protective activities of the filamentous hemagglutinin and the lymphocytosis-promoting factor of Bordetella pertussis in mice. , 1984, The Journal of infectious diseases.

[29]  E N Trifonov,et al.  A computer algorithm for testing potential prokaryotic terminators. , 1984, Nucleic acids research.

[30]  J. Devereux,et al.  A comprehensive set of sequence analysis programs for the VAX , 1984, Nucleic Acids Res..

[31]  M. Gribskov,et al.  The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression , 1984, Nucleic Acids Res..

[32]  E. Hewlett,et al.  Tn5-induced mutations affecting virulence factors of Bordetella pertussis , 1983, Infection and immunity.

[33]  L. Irons,et al.  Heterogeneity of the filamentous haemagglutinin of Bordetella pertussis studied with monoclonal antibodies. , 1983, Journal of general microbiology.

[34]  H. Sato,et al.  Separation and purification of the hemagglutinins from Bordetella pertussis , 1983, Infection and immunity.

[35]  R. Isaacson,et al.  Proteinaceous bacterial adhesins and their receptors. , 1983, Critical reviews in microbiology.

[36]  J. Fickett.  Recognition of protein coding regions in DNA sequences. , 1982, Nucleic acids research.

[37]  A. M. Collier,et al.  Mycoplasma pneumoniae infection: role of a surface protein in the attachment organelle. , 1982, Science.

[38]  W. Fiers,et al.  Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. , 1981, Gene.

[39]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[40]  D Court,et al.  Regulatory sequences involved in the promotion and termination of RNA transcription. , 1979, Annual review of genetics.

[41]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[42]  P. Sharp,et al.  Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids , 1977, Cell.

[43]  N. Davidson,et al.  Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. , 1977, Nucleic acids research.

[44]  A. M. Pappenheimer.  Diphtheria toxin. , 1952, Annual review of biochemistry.

[45]  K. Drlica,et al.  Conformational transitions in the Escherichia coli chromosome: analysis by viscometry and sedimentation. , 1975, Journal of molecular biology.

[46]  J. Shine,et al.  The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. , 1974, Proceedings of the National Academy of Sciences of the United States of America.