Inactivation of the gene encoding surface protein SspA in Streptococcus gordonii DL1 affects cell interactions with human salivary agglutinin and oral actinomyces.
Cell surface protein SSP-5 in the oral bacterium Streptococcus gordonii M5 binds human salivary agglutinin in a Ca(2+)-dependent reaction (D.R. Demuth, E.E. Golub, and D. Malamud, J. Biol. Chem. 265:7120-7126, 1990). The region of the gene encoding an N-terminal segment of a related polypeptide (SspA) in S. gordonii DL1 (Challis) was isolated following polymerase chain reaction amplification of genomic DNA. The sspA gene in S. gordonii DL1 was insertionally inactivated by homologous recombination of the erythromycin resistance (Emr) determinant ermAM onto the streptococcal chromosome. The SspA polypeptide (apparent molecular mass, 210 kDa) was detected on Western blots (immunoblots) of spheroplast extracts and extracellular culture medium proteins from wild-type strain DL1 but was absent from Emr mutants. One SspA- mutant (designated OB220) was not altered in rate or extent of aggregation by whole saliva or parotid saliva but showed reduced aggregation in the presence of purified salivary agglutinin. Mutant bacteria were unaffected in their ability to adhere to hydroxylapatite beads coated with whole or parotid saliva and were unaltered in cell surface hydrophobicity. However, the SspA- strain OB220 was deficient in binding salivary agglutinin and in binding to six strains of Actinomyces naeslundii. Therefore, expression of SspA polypeptide in S. gordonii is associated with both agglutinin-dependent and agglutinin-independent aggregation and adherence reactions of streptococcal cells.
Jenkinson
H F
HF
Department of Oral Biology and Oral Pathology, University of Otago, Dunedin, New Zealand.
Terry
S D
SD
McNab
R
R
Tannock
G W
GW
D000190:Actinomyces / Q000502:physiology*; D018829:Adhesins, Bacterial; D000373:Agglutinins / Q000494:pharmacology*; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002449:Cell Aggregation; D005798:Genes, Bacterial; D005964:Glucosyltransferases / Q000096:biosynthesis; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009055:Mouth / Q000382:microbiology*; D009154:Mutation; D012463:Saliva / Q000502:physiology*; D013291:Streptococcus / Q000235:genetics* / Q000502:physiology
D018829:Adhesins, Bacterial; D000373:Agglutinins; D001426:Bacterial Proteins; D008565:Membrane Proteins; C082314:SspA protein, bacteria; D005964:Glucosyltransferases