Role of the major pneumococcal autolysin in the atypical response of a clinical isolate of Streptococcus pneumoniae.
The autolytic enzyme (an N-acetylmuramyl-L-alanine amidase) of a clinical isolate, strain 101/87, which is classified as an atypical pneumococcus, has been studied for the first time. The lytA101 gene coding for this amidase (LYTA101) has been cloned, sequenced, and expressed in Escherichia coli. The LYTA101 amidase has been purified and shown to be similar to the main autolytic enzyme (LYTA) present in the wild-type strain of Streptococcus pneumoniae, although it exhibits a lower specific activity, a higher sensitivity to inhibition by free choline, and a modified thermosensitivity with respect to LYTA. Most important, in contrast with the LYTA amidase, the activity of the LYTA101 amidase was inhibited by sodium deoxycholate. This property is most probably responsible of the deoxycholate-insensitive phenotype shown by strain 101/87. Phenotypic curing of strain 101/87 by externally adding purified LYTA or LYTA101 amidase restored in this strain some typical characteristics of the wild-type strain of pneumococcus (e.g., formation of diplo cells and sensitization to lysis by sodium deoxycholate), although the amount of the LYTA101 amidase required to restore these properties was much higher than in the case of the LYTA amidase. Our results indicate that modifications in the primary structure or in the mechanisms that control the activity of cell wall lytic enzymes seem to be responsible for the characteristics exhibited by some strains of S. pneumoniae that have been classically misclassified and should be now considered atypical pneumococcal strains.
Díaz
E
E
Unidad de Genética Bacteriana, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
López
R
R
García
J L
JL
D000581:Amidohydrolases / Q000737:chemistry; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D003840:Deoxycholic Acid / Q000494:pharmacology; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry* / Q000235:genetics; D015183:Restriction Mapping; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D003840:Deoxycholic Acid; D000581:Amidohydrolases; D009238:N-Acetylmuramoyl-L-alanine Amidase; C019665:amidase