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. 1982 Nov;152(2):722–735. doi: 10.1128/jb.152.2.722-735.1982

Replication and incompatibility properties of plasmid pE194 in Bacillus subtilis.

T J Gryczan, J Hahn, S Contente, D Dubnau
PMCID: PMC221522  PMID: 6290448

Abstract

pE194, a 3.5-kilobase multicopy plasmid, confers resistance to the macrolide-lincosamide-streptogramin B antibiotics in Bacillus subtilis. By molecular cloning and deletion analysis we have identified a replication segment on the physical map of this plasmid, which consists of about 900 to 1,000 base pairs. This segment contains the replication origin. It also specifies a trans-acting function (rep) required for the stable replication of pE194 and a negatively acting copy control function which is the product of the cop gene. The target sites for the rep and cop gene products are also within this region. Two incompatibility determinants have been mapped on the pE194 genome and their properties are described. One (incA) resides within the replication region and may be identical to cop. incB, not located in the replication region, expresses incompatibility toward a copy control mutant (cop-6) but not toward the wild-type replicon.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  2. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  3. Contente S., Dubnau D. Characterization of plasmid transformation in Bacillus subtilis: kinetic properties and the effect of DNA conformation. Mol Gen Genet. 1979 Jan 2;167(3):251–258. doi: 10.1007/BF00267416. [DOI] [PubMed] [Google Scholar]
  4. Contente S., Dubnau D. Marker rescue transformation by linear plasmid DNA in Bacillus subtilis. Plasmid. 1979 Oct;2(4):555–571. doi: 10.1016/0147-619x(79)90054-4. [DOI] [PubMed] [Google Scholar]
  5. Donoghue D. J., Sharp P. A. Replication of colicin E1 plasmid DNA in vivo requires no plasmid-encoded proteins. J Bacteriol. 1978 Mar;133(3):1287–1294. doi: 10.1128/jb.133.3.1287-1294.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubnau D., Davidoff-Abelson R. Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol. 1971 Mar 14;56(2):209–221. doi: 10.1016/0022-2836(71)90460-8. [DOI] [PubMed] [Google Scholar]
  7. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gryczan T. J., Dubnau D. Construction and properties of chimeric plasmids in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1428–1432. doi: 10.1073/pnas.75.3.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gryczan T. J., Grandi G., Hahn J., Grandi R., Dubnau D. Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance. Nucleic Acids Res. 1980 Dec 20;8(24):6081–6097. doi: 10.1093/nar/8.24.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gryczan T., Shivakumar A. G., Dubnau D. Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis. J Bacteriol. 1980 Jan;141(1):246–253. doi: 10.1128/jb.141.1.246-253.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982 May;150(2):804–814. doi: 10.1128/jb.150.2.804-814.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horinouchi S., Weisblum B. Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7079–7083. doi: 10.1073/pnas.77.12.7079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Horinouchi S., Weisblum B. The control region for erythromycin resistance: free energy changes related to induction and mutation to constitutive expression. Mol Gen Genet. 1981;182(2):341–348. doi: 10.1007/BF00269681. [DOI] [PubMed] [Google Scholar]
  14. Inuzuka M., Helinski D. R. Requirement of a plasmid-encoded protein for replication in vitro of plasmid R6K. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5381–5385. doi: 10.1073/pnas.75.11.5381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iordănescu S. Three distinct plasmids originating in the same Staphylococcus aureus strain. Arch Roum Pathol Exp Microbiol. 1976 Jan-Jun;35(1-2):111–118. [PubMed] [Google Scholar]
  16. Ishii K., Hashimoto-Gotoh T., Matsubara K. Random replication and random assortment model for plasmid incompatibility in bacteria. Plasmid. 1978 Sep;1(4):435–445. doi: 10.1016/0147-619x(78)90002-1. [DOI] [PubMed] [Google Scholar]
  17. Itoh T., Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proc Natl Acad Sci U S A. 1980 May;77(5):2450–2454. doi: 10.1073/pnas.77.5.2450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Khan S. A., Carleton S. M., Novick R. P. Replication of plasmid pT181 DNA in vitro: requirement for a plasmid-encoded product. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4902–4906. doi: 10.1073/pnas.78.8.4902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kline B. C. Incompatibility between Flac, R386, and F:pSC101 recombinant plasmids: the specificity of F incompatibility genes. Plasmid. 1979 Jul;2(3):437–445. doi: 10.1016/0147-619x(79)90027-1. [DOI] [PubMed] [Google Scholar]
  20. Lovett M. A., Katz L., Helinski D. R. Unidirectional replication of plasmid ColE1 DNA. Nature. 1974 Sep 27;251(5473):337–340. doi: 10.1038/251337a0. [DOI] [PubMed] [Google Scholar]
  21. Lovett M. A., Sparks R. B., Helinski D. R. Bidirectional replication of plasmid R6K DNA in Escherichia coli; correspondence between origin of replication and position of single-strand break in relaxed complex. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2905–2909. doi: 10.1073/pnas.72.8.2905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morell P., Smith I., Dubnau D., Marmur J. Isolation and characterization of low molecular weight ribonucleic acid species from Bacillus subtilis. Biochemistry. 1967 Jan;6(1):258–265. doi: 10.1021/bi00853a040. [DOI] [PubMed] [Google Scholar]
  23. Novick R. P., Adler G. K., Majumder S., Khan S. A., Carleton S., Rosenblum W. D., Iordanescu S. Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4108–4112. doi: 10.1073/pnas.79.13.4108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Novick R. P., Hoppensteadt F. C. On plasmid incompatibility. Plasmid. 1978 Sep;1(4):421–434. doi: 10.1016/0147-619x(78)90001-x. [DOI] [PubMed] [Google Scholar]
  25. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Scheer-Abramowitz J., Gryczan T. J., Dubnau D. Origin and mode of replication of plasmids pE194 and pUB110. Plasmid. 1981 Jul;6(1):67–77. doi: 10.1016/0147-619x(81)90054-8. [DOI] [PubMed] [Google Scholar]
  27. Shepard H. M., Gelfand D. H., Polisky B. Analysis of a recessive plasmid copy number mutant: evidence for negative control of Col E1 replication. Cell. 1979 Oct;18(2):267–275. doi: 10.1016/0092-8674(79)90046-1. [DOI] [PubMed] [Google Scholar]
  28. Shivakumar A. G., Dubnau D. Characterization of a plasmid-specified ribosome methylase associated with macrolide resistance. Nucleic Acids Res. 1981 Jun 11;9(11):2549–2562. doi: 10.1093/nar/9.11.2549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shivakumar A. G., Gryczan T. J., Kozlov Y. I., Dubnau D. Organization of the pE194 genome. Mol Gen Genet. 1980;179(2):241–252. doi: 10.1007/BF00425450. [DOI] [PubMed] [Google Scholar]
  30. Shivakumar A. G., Hahn J., Grandi G., Kozlov Y., Dubnau D. Posttranscriptional regulation of an erythromycin resistance protein specified by plasmic pE194. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3903–3907. doi: 10.1073/pnas.77.7.3903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Timmis K. N., Danbara H., Brady G., Lurz R. Inheritance functions of group IncFII transmissible antibiotic resistance plasmids. Plasmid. 1981 Jan;5(1):53–75. doi: 10.1016/0147-619x(81)90077-9. [DOI] [PubMed] [Google Scholar]
  32. Veltkamp E., Stuitje A. R. Replication and structure of the bacteriocinogenic plasmids Clo DF13 and CoI E1. Plasmid. 1981 Jan;5(1):76–99. doi: 10.1016/0147-619x(81)90078-0. [DOI] [PubMed] [Google Scholar]
  33. Weisblum B., Graham M. Y., Gryczan T., Dubnau D. Plasmid copy number control: isolation and characterization of high-copy-number mutants of plasmid pE194. J Bacteriol. 1979 Jan;137(1):635–643. doi: 10.1128/jb.137.1.635-643.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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