Nucleotide sequence of the cell wall proteinase gene of Streptococcus cremoris Wg2.
A 6.5-kilobase HindIII fragment that specifies the proteolytic activity of Streptococcus cremoris Wg2 was sequenced entirely. The nucleotide sequence revealed two open reading frames (ORFs), a small ORF1 with 295 codons and a large ORF2 containing 1,772 codons. For both ORFs, there was no stop codon on the HindIII fragment. A partially overlapping PstI fragment was used to locate the translation stop of the large ORF2. The entire ORF2 contained 1,902 coding triplets, followed by an apparently rho-independent terminator sequence. The inferred amino acid sequence would result in a protein of 200 kilodaltons. Both ORFs have their putative transcription and translation signals in a 345-base-pair ClaI fragment. ORF2 is preceded by a promoter region containing a 15-base-pair complementary direct repeat. Both the truncated 33- and the 200-kilodalton proteins have a signal peptide-like N-terminal amino acid sequence. The protein specified by ORF2 contained regions of extensive homology with serine proteases of the subtilisin family. Specifically, amino acid sequences involved in the formation of the active site (viz., Asp-32, His-64, and Ser-221 of the subtilisins) are well conserved in the S. cremoris Wg2 proteinase. The homologous sequences are separated by nonhomologous regions which contain several inserts, most notably a sequence of approximately 200 amino acids between the His and Ser residues of the active site.
Kok
J
J
Institute of Genetics, University of Groningen, Haren, The Netherlands.
Leenhouts
K J
KJ
Haandrikman
A J
AJ
Ledeboer
A M
AM
Venema
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002473:Cell Wall / Q000201:enzymology; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial / Q000032:analysis; D010450:Endopeptidases / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000032:analysis; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics; D013381:Subtilisins / Q000032:analysis / Q000235:genetics
D003062:Codon; D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D010450:Endopeptidases; D013381:Subtilisins