title: A comprehensive genetic study of streptococcal immunoglobulin A1 proteases: evidence for recombination within and between species.
abstract: An analysis of 13 immunoglobulin A1 (IgA1) protease genes (iga) of strains of Streptococcus pneumoniae, Streptococcus oralis, Streptococcus mitis, and Streptococcus sanguis was carried out to obtain information on the structure, polymorphism, and phylogeny of this specific protease, which enables bacteria to evade functions of the predominant Ig isotype on mucosal surfaces. The analysis included cloning and sequencing of iga genes from S. oralis and S. mitis biovar 1, sequencing of an additional seven iga genes from S. sanguis biovars 1 through 4, and restriction fragment length polymorphism (RFLP) analyses of iga genes of another 10 strains of S. mitis biovar 1 and 6 strains of S. oralis. All 13 genes sequenced had the potential of encoding proteins with molecular masses of approximately 200 kDa containing the sequence motif HEMTH and an E residue 20 amino acids downstream, which are characteristic of Zn metalloproteinases. In addition, all had a typical gram-positive cell wall anchor motif, LPNTG, which, in contrast to such motifs in other known streptococcal and staphylococcal proteins, was located in their N-terminal parts. Repeat structures showing variation in number and sequence were present in all strains and may be of relevance to the immunogenicities of the enzymes. Protease activities in cultures of the streptococcal strains were associated with species of different molecular masses ranging from 130 to 200 kDa, suggesting posttranslational processing possibly as a result of autoproteolysis at post-proline peptide bonds in the N-terminal parts of the molecules. Comparison of deduced amino acid sequences revealed a 94% similarity between S. oralis and S. mitis IgA1 proteases and a 75 to 79% similarity between IgA1 proteases of these species and those of S. pneumoniae and S. sanguis, respectively. Combined with the results of RFLP analyses using different iga gene fragments as probes, the results of nucleotide sequence comparisons provide evidence of horizontal transfer of iga gene sequences among individual strains of S. sanguis as well as among S. mitis and the two species S. pneumoniae and S. oralis. While iga genes of S. sanguis and S. oralis were highly homogeneous, the genes of S. pneumoniae and S. mitis showed extensive polymorphism reflected in different degrees of antigenic diversity.
authors.affiliation: Department of Medical Microbiology, University of Aarhus, Denmark. mikrkp@svfcd.aau.dk
authors.forename: K
authors.initials: K
authors.lastname: Poulsen
authors.forename: J
authors.initials: J
authors.lastname: Reinholdt
authors.forename: C
authors.initials: C
authors.lastname: Jespersgaard
authors.forename: K
authors.initials: K
authors.lastname: Boye
authors.forename: T A
authors.initials: TA
authors.lastname: Brown
authors.forename: M
authors.initials: M
authors.lastname: Hauge
authors.forename: M
authors.initials: M
authors.lastname: Kilian
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D015723:Gene Library; D016680:Genome, Bacterial; D018928:Immunity, Mucosal; D008666:Metalloendopeptidases / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D011499:Protein Processing, Post-Translational; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase; D008666:Metalloendopeptidases