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Genetic organization of the streptokinase region of the Streptococcus equisimilis H46A chromosome

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Abstract

The complete nucleotide sequences of four genes and one open reading frame (ORF1) adjacent to the streptokinase gene, skc, from Streptococcus equisimilis H46A were determined. These genes are encoded on the opposite DNA strand to skc and are arranged as follows: dexB-abc-lrp-skc-ORF1-rel. The dexB gene, coding for an α-glucosidase (Mr 61733), and abc, encoding an ABC transporter (Mr 42080), are similar to the dexB and msmK genes, respectively, from the multiple sugar metabolism operon of S. mutans. The lrp gene specifies a leucine-rich protein (Mr 32302) that has a leucine-zipper motif at its C-terminus. The function of the Lrp protein is not known but appeared to be detrimental when overexpressed in Escherichia coli. Although lrp appears not to be an essential gene, as judged by plasmid insertion mutagenesis, it is conserved in all streptococcal strains carrying a streptokinase gene. The rel gene showed significant homology to the E. coli relA and spoT genes involved in the stringent response to amino acid deprivation. Multiple alignment of the amino acid sequences of Rel (Mr 83913), RelA and SpoT revealed 59.4% homology of the primary structures. Northern hybridiziatioo analyses of the genes in the skc region showed skc to be transcribed most abundantly. In addition to transcripts for skc, monocistronic mRNAs were detected for all three genes divergently transcribed from skc. Although there was also some read-through transcription from lrp into abc, and from abc into dexB, the transcription pattern suggests a high degree of transcriptional and functional independence not only of skc but also abc and dexB. Prominent structural features in intergenic regions included a static DNA bending locus located upstream and a putative bidirectional transcription terminator downstream of skc.

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References

  • Abel T, Maniatis T (1989) Action of leucine zippers. Nature 341:24–25

    Google Scholar 

  • Alloing G, Trombe M-C, Claverys J-P (1990) The ami locus of the gram-positive bacterium Streptococcus pneumoniae is similar to binding protein-dependent transport operons of gram-negative bacteria. Mol Microbiol 4:633–644

    Google Scholar 

  • Caparon MG, Scott JR (1991) Genetic manipulation of pathogenic streptococci. Methods Enzymol 204:556–586

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Christensen LR (1945) Streptococcal fibrinolysin: a proteolytic reaction due to a serum enzyme activated by streptococcal fibrinolysin. J Gen Physiol 28:363–383

    Google Scholar 

  • Dahl MK, Francoz E, Saurin W, Boos W, Manson MD, Hofnung M (1989) Comparison of sequences from the malB regions of Salmonella typhimurium and Enterobacter aerogenes with Escherichia coli K12: a potential new regulatory site in the interoperonic region. Mol Gen Genet 218:199–207

    Google Scholar 

  • Feng DF, Doolittle RF (1987) Progressive sequence alignment as a prerequisite to correct phylogenetic trees. J Mol Evol 25:351–360

    Google Scholar 

  • Galli D, Friesenegger A, Wirth R (1992) Transcriptional control of sex-pheromone-inducible genes on plasmid pAD1 of Enterococcus faecalis and sequence analysis of a third structural gene for (pPDI-encoded) agregation substance. Mol Microbiol 6:1297–1308

    Google Scholar 

  • GISSI (1990) GISSI-2: a factorial randomised trial of alteplase versus streptokinase and heparin versus no heparin among 12 490 patients with acute myocardial infarction. Lancet 336:65–71

    Google Scholar 

  • Henco K (1991) The QIAexpressionist. Diagen, Düsseldorf

    Google Scholar 

  • Higgins CF (1992) ABC transporters: from microorganisms to man. Annu Rev Cell Biol 8:67–113

    Google Scholar 

  • Higgins CF, Haag PD, Nikaido K, Ardeshir F, Garcia G, Ames GF-L (1982) Complete nucleotide sequence and identification of membrane components of the histidine transport operon of S. typhimurium. Nature 298:723–727

    Google Scholar 

  • Hiles ID, Gallagher MP, Jamieson DJ, Higgins CF (1987) Molecular characterization of the oligopeptide permease of Salmonella typhimurium. J Mol Biol 195:125–142

    Google Scholar 

  • Holm SE (1988) The pathogenesis of acute post-streptococcal glomerulonephritis in new lights. Acta Pathol Microbiol Immunol Scand 96:189–193

    Google Scholar 

  • Huang T-T, Malke H, Ferretti JJ (1989a) Heterogeneity of the streptokinase gene in group A streptococci. Infect Immun 57:502–506

    Google Scholar 

  • Huang T-T, Malke H, Ferretti JJ (1989b) The streptokinase gene of group A streptococci: cloning, expression in Escherichia coli, and sequence analysis. Mol Microbiol 3:197–205

    Google Scholar 

  • Johnston KH, Chaiban JE, Wheeler RC (1991) Analysis of the variable domain of the streptokinase gene from group A streptococci by the polymerase chain reaction. In: Dunny GM, Cleary PP, McKay LL (eds) Genetics and molecular biology of streptococci, lactococci, and enterococci. American Society of Microbiology, Washington DC, pp 190–194

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lipman DJ, Pearson WR (1985) Rapid and sensitive similarity searches. Science 227:1435–1441

    Google Scholar 

  • Lottenberg R, DesJardin LE, Wang H, Boyle MDP (1992) Streptokinase-producing streptococci grown in human plasma acquire unregulated cell-associated plasmin activity. J Infect Dis 166:436–440

    Google Scholar 

  • Macrina FL, Evans RP, Tobian JA, Hartley DL, Clewell DB, Jones KR (1983) Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. Gene 25:145–150

    Google Scholar 

  • Malke H (1993) Polymorphism of the streptokinase gene: implications for the pathogenesis of post-streptococcal glomerulonephritis. Zentralbl Bakteriol 278:246–257

    Google Scholar 

  • Malke H, Ferretti JJ (1984) Streptokinase: cloning, expression, and excretion by Escherichia coli. Proc Natl Acad Sci USA 81:3557–3561

    Google Scholar 

  • Malke H, Roe B, Ferretti JJ (1985) Nucleotide sequence of the streptokinase gene from Streptococcus equisimilis H46A. Gene 34:357–362

    Google Scholar 

  • McDowell TD, Reed KE, Harley WM (1988) Accumulation of ppGpp in three streptococci during periods of amino acid starvation. FEMS Microbiol Lett 56:151–156

    Google Scholar 

  • McMaster GK, Carmichael GG (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74:4835–4838

    Google Scholar 

  • Metzger S, Dror IB, Aizenman E, Schreiber G, Toone M, Friesen JD, Cashel M, Glaser G (1988) The nucleotide sequence and characterization of the relA gene of Escherichia coli. J Biol Chem 263:15699–15704

    Google Scholar 

  • Metzger S, Sarubbi E, Glaser G, Cashel M (1989) Protein sequences encoded by the relA and the spoT genes of Escherichia coli are interrelated. J Biol Chem 264:9122–9125

    Google Scholar 

  • Müller J, Malke H (1989) Sequence-directed DNA bending upstream of the streptokinase promoter. J Basic Microbiol 29:611–616

    Google Scholar 

  • Ochman H, Wilson AC (1987) Evolution in bacteria: evidence for a universal substitution rate in cellular genomes. J Mol Evol 26:74–86

    Google Scholar 

  • Ohkuni H, Todome Y, Suzuki H, Mizuse M, Kotani N, Horiuchi K, Shikama N, Tsugita A, Johnston KH (1992) Immunochemical studies and complete amino acid sequence of the streptokinase gene from Streptococcus pyogenes (group A) M type 12 strain A374. Infect Immun 60:278–283

    Google Scholar 

  • Overduin P, Boss W, Tommassen J (1988) Nucleotide sequence of the ugp genes of E. coli K-12: homology to the maltose system. Mol Microbiol 2:767–775

    Google Scholar 

  • Perego M, Higgins CF, Pearce SR, Gallagher MP, Hoch JA (1991) The oligopeptide transport system of Bacillus subtilis plays a role in the initiation of sporulation. Mol Microbiol 5:173–185

    Google Scholar 

  • Plaskon RR, Wartell RM (1987) Sequence distributions associated with DNA curvature are found upstream of strong E. coli promoters. Nucleic Acids Res 15:785–795

    Google Scholar 

  • Russell RRB, Ferretti JJ (1990) Nucleotide sequence of the dextran glucosidase (dexB) gene of Streptococcus mutans. J Gen Microbiol 136:803–810

    Google Scholar 

  • Russell RRB, Aduse-Opoku J, Sutcliffe IC, Tao L, Ferretti JJ (1992) A binding protein-dependent transport system in Streptococcus mutans responsible for multiple sugar metabolism. J Biol Chem 267:4631–4637

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis TE (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693

    Google Scholar 

  • Sarubbi E, Rudd KE, Xiao H, Ikehara K, Kalman M, Cashel M (1989) Characterization of the spoT gene of Escherichia coli. J Biol Chem 264:15074–15082

    Google Scholar 

  • Scheidtmann KH (1990) Immunological detection of proteins of known sequence. In: Creighton TE (ed) Protein structure: a practical approach. IRL Press, Oxford, pp 93–115

    Google Scholar 

  • Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    Google Scholar 

  • Vinson CR, Sigler PB, McKnight SL (1989) Scissors-grip model for DNA recognition by a family of leucine zipper proteins. Science 246:911–916

    Google Scholar 

  • Walter F, Siegel M, Malke H (1989a) Nucleotide sequence of the streptokinase gene from a Streptococcus pyogenes type 1 strain. Nucleic Acids Res 17:1261

    Google Scholar 

  • Walter F, Siegel M, Malke H (1989b) Nucleotide sequence of the streptokinase gene from a group-G Streptococcus. Nucleic Acids Res 17:1262

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33:103–119

    Google Scholar 

  • Zamenhof PJ, Villarejo M (1972) Construction and properties of Escherichia coli strains exhibiting α-complementation of β-ga-lactosidase fragments in vivo. J Bacteriol 110:171–178

    Google Scholar 

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Communicated by H. Böhme

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Mechold, U., Steiner, K., Vettermann, S. et al. Genetic organization of the streptokinase region of the Streptococcus equisimilis H46A chromosome. Molec. Gen. Genet. 241, 129–140 (1993). https://doi.org/10.1007/BF00280210

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  • DOI: https://doi.org/10.1007/BF00280210

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