Molecular characterization of group A streptococcal (GAS) oligopeptide permease (opp) and its effect on cysteine protease production.
Bacterial oligopeptide permeases are membrane-associated complexes of five proteins belonging to the ABC-transporter family, which have been found to be involved in obtaining nutrients, cell-wall metabolism, competence, and adherence to host cells. A lambda library of the strain CS101 group A streptococcal (GAS) genome was used to sequence 10,192 bp containing the five genes oppA to oppF of the GAS opp operon. The deduced amino acid sequences exhibited 50-84% homology to pneumococcal AmiA to AmiF sequences. The operon organization of the five genes was confirmed by transcriptional analysis and an additional shorter oppA transcript was detected. Insertional inactivation was used to create serotype M49 strains which did not express either the oppA gene or the ATPase genes, oppD and oppF. The mutation in oppA confirmed that the additional shorter oppA transcript originated from the opp operon and was probably due to an intra-operon transcription terminator site located downstream of oppA. While growth kinetics, binding of serum proteins, and attachment to eukaryotic cells were unaffected, the oppD/F mutants showed reduced production of the cysteine protease, SpeB, and a change in the pattern of secreted proteins. Thus, the GAS opp operon appears to contribute to both protease production and export/processing of secreted proteins.
Podbielski
A
A
Institute of Medical Microbiology, Hospital of the Technical University (Klinikum RWTH), Aachen, Germany. andreasp@imib.rwth-aachen.de
Pohl
B
B
Woischnik
M
M
Körner
C
C
Schmidt
K H
KH
Rozdzinski
E
E
Leonard
B A
BA
D000595:Amino Acid Sequence; D000630:Aminopterin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins; D001483:Base Sequence; D001692:Biological Transport; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004352:Drug Resistance, Microbial; D026901:Membrane Transport Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D009876:Operon; D009943:Organophosphorus Compounds / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013297:Streptococcus pyogenes / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D009943:Organophosphorus Compounds; C003122:bialaphos; C037230:oligopeptide permease, Bacteria; D003546:Cysteine Endopeptidases; C042477:streptopain; D000630:Aminopterin