Summary
Insertion of the erythromycin resistance transposon Tn551 into a single site of the Staphylococcus aureus chromosome resulted in decreased production of alphatoxin, serine and metallo-proteinases and several other extracellular proteins and a simultaneous increase in the production of protein A. The site of insertion, designated exp, was separate from the structural gene for alpha-toxin and protein A. Hybridization analysis showed that the effect of the insertional mutation on the expression of the alphatoxin and protein A was at the level of transcription. The chromosomal DNA flanking the transposon and the corresponding DNA of the wild-type strain was cloned in Escherichia coli. Northern blot hybridization experiments revealed that the exp locus codes for a major RNA of approximately 3.5 kb. This RNA was not found in the insertional mutant nor in a spontaneous exp mutant. A map of the exp locus constructed by Northern blot and restriction enzyme analysis showed that the insertional mutation was located in the middle of the coding sequence of the 3.5 kb RNA. The insertional mutant was reverted to wild type by inserting a recombinant plasmid containing most of the coding sequence of the 3.5 kb RNA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abbas-Ali B, Coleman G (1977) the characteristics of extracellular protein secretion by Staphylococcus aureus (Wood 46) and their relationships to the regulation of alpha-toxin. J Gen Microbiol 92:277–282
Barnes WM (1977) Plasmid detection and sizing in single colony lysates. Science 195:393–394
Berger-Bäche B (1983) Insertional inactivation of staphylococcal methicillin resistance by Tn551. J Bacteriol 154:479–487
Björklind A, Arvidson S (1980) Mutants of Staphylococcus aureus affected in the regulation of exoprotein synthesis. FEMS Microbiol Lett 7:203–206
Bolivar F, Rodriges RL, Grene PY, Betlach MC, Heynecker HL, Boyer HW, Crosa YH, Falkow S (1977) Construction and characterization of new cloning vehicles, II. A multipurpose cloning system. Gene 2:95–113
Botchan M, Topo W, Sambrook J (1976) The arrangement of Simian virus 40 sequences in the DNA of transformed cells. Cell 9:269–287
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Coleman G (1981) Pleiotropic compensation in the regulation of extracellular protein formation by a low alpha-toxin-producing variant of Staphylococcus aureus (Wood 46). J Gen Microbiol 122:11–15
Coleman G, Abbas-Ali B (1977) Comparison of the patterns of increase in alpha-toxin and total extracellular protein by Staphylococcus aureus (Wood 46) grown in media supproting widely differing growth characteristics. Infect Immun 17:278–281
Grunstein M, Hogness DS (1975) Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72:3961–3965
Hattori M, Sakaki Y (1986) Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 152:232–238
Iordanescu S (1975) Recombinant plasmid obtained from two different compatible staphylococcal plasmids. J Bacteriol 124:597–601
Janzon L, Löfdahl S, Arvidson S (1986) Evidence for a coordinate transcriptional control of alpha-toxin and protein A in Staphylococcus aureus. FEMS Microbiol Lett 33:193–198
Khan SA, Novick RP (1980) Terminal nucleotide sequences of Tn551, a transposon specifying erythromycin resistance in Staphylococcus aureus: Homology with Tn3. Plasmid 4:148–154
Kehoe M, Duncan J, Foster T, Fairweather N, Dougan G (1983) Cloning, expression and mapping of the Staphylococcus aureus α-hemolysin determinant in Escherichia coli K-12. Infect Immun 41:1105–1111
Lindberg M (1981) Genetic studies in Staphylococcus aureus using protoplasts: Cell fusion and transformation. In: Jeljaszewicz J (ed) Staphylococci and staphylococcal infections. Gustav Fischer, Stutgart, pp 534–541
Löfdahl S, Guss B, Uhlén M, Philipson L, Lindberg M (1983) Gene for staphylococcal protein A. Proc Natl Acad Sci USA 80:697–701
Meinkoth J, Wahl G (1984) Hybridization of nucleic acids immobilized on solid supports. Anal Biochem 138:267–284
Melton DA, Krieg J, Rebagliati MR, Maniatis T, Zinn K, Green M (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056
Novick RP (1967) Properties of a cryptic high frequency transducing phage in Staphylococcus aureus. Virology 33:155–166
Novick RP (1974) Studies on plasmid replication, III. Isolation and characterisation of replication defective mutants. Mol Gen Genet 135:131–147
Novick RP, Murphy E, Gryczan TJ, Baron E, Edelman I (1979a) Penicillinase plasmids of Staphylococcus aureus: Restriction deletion maps. Plasmid 2:109–129
Novick RP, Edelman I, Schweisinger MD, Gruss AD, Swanson EC, Pattee PA (1979b) Genetic translocation in Staphylococcus aureus. Proc Natl Acad Sci USA 76:400–404
Pattee PA (1981) Distribution of Tn551 insertion sites responsible for auxotrophy on the Staphylococcus aureus chromosome. J Bacteriol 145:479–488
Recsei P, Kreiswirth B, O'Reilly M, Schlivert R, Gruss A, Novick RP (1986) Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Mol Gen Genet 202:58–61
Smith JM (1979) Staphylococcus aureus. In: Mandell GL, Douglas R, Bennet J (eds) Principles and practice of infectious diseases. Wiley and Son. New York, pp 1530–1552
Yoshikawa M, Matsudo F, Naka M, Murofuski E, Tsunematsu Y (1974) Pleiotropic alteration of activities of several toxins and enzymes in mutants of Staphylococcus aureus. J Bacteriol 119:143–148
Author information
Authors and Affiliations
Additional information
Communicated W. Goebel
Rights and permissions
About this article
Cite this article
Morfeldt, E., Janzou, L., Arvidson, S. et al. Cloning of a chromosomal locus (exp) which regulates the expression of several exoprotein genes in Staphylococcus aureus . Molec. Gen. Genet. 211, 435–440 (1988). https://doi.org/10.1007/BF00425697
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00425697