Structural gene and strain specificity of a novel cysteine protease produced by Staphylococcus aureus isolated from a diseased chicken

• Published in: Veterinary microbiology 2002-10, Vol.89 (2), p.201-210
• Main Authors: Takeuchi, Shotaro, Matsunaga, Kazue, Inubushi, Sachiko, Higuchi, Hiroki, Imaizumi, Keisuke, Kaidoh, Toshio
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Avian strain; Bacteriology; Base Sequence; Biological and medical sciences; Blotting, Southern - veterinary; Blotting, Western - veterinary; Chickens; Cloning, Molecular; Cysteine Endopeptidases - genetics; Cysteine Endopeptidases - isolation & purification; Cysteine Endopeptidases - metabolism; Cysteine protease; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genetics; Microbiology; Molecular Sequence Data; Molecular Weight; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Polymerase Chain Reaction - veterinary; Poultry Diseases - metabolism; Poultry Diseases - microbiology; Recombinant Proteins - genetics; ScpA gene; Sequence Homology, Amino Acid; Species Specificity; Staphylococcal Infections - metabolism; Staphylococcal Infections - microbiology; Staphylococcal Infections - veterinary; Staphylococcus aureus; Staphylococcus aureus - enzymology; Staphylococcus aureus - genetics
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/S0378-1135(02)00171-2

Recently whole genome sequencing of Staphylococcus aureus has revealed the genes encoding cysteine proteases such as staphopain and SspB. In this study, we cloned and sequenced the structural gene ( ScpA) encoding a cysteine (thiol) protease of S. aureus strain CH-91 from a chicken with dermatitis using polymerase chain reaction (PCR) and inverse PCR methods. The sequence information revealed a coding sequence (CDS) of 1200 nucleotides encoding the ScpA preproenzyme of 399 amino acids with a molecular mass of 45,071 Da. The deduced amino acid sequence of the ScpA differed at many positions from those of staphopain and SspB with identities of 64 and 42%, respectively. In the Southern blot analysis with a total DNA of S. aureus strain CH-91, the ScpA probe hybridized with a single 7.7 kb XbaI fragment or 2.8 and 0.8 kb EcoRI fragments, whereas the staphopain and SspB probes did not hybridize with these DNA fragments. These results suggest that this ScpA gene is a single-copy gene and is a novel gene, which is not found in the published whole genome sequences of S. aureus. In immunoblot, PCR, and Southern blot assays, the ScpA or its gene was detected in high protease-producing strains from chickens, but was not recognized in bovine and porcine strains or low protease-producing avian strains. These results indicate that the ScpA of CH-91 type may be specific to the high protease-producing strains of S. aureus from chickens, namely, there is a strain specificity of the ScpA.