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Molecular and General Genetics MGG

, Volume 241, Issue 1–2, pp 97–105 | Cite as

Replication of the promiscuous plasmid pLSI: a region encompassing the minus origin of replication is associated with stable plasmid inheritance

  • Gloria del Solar
  • Gabriela Kramer
  • Sara Ballester
  • Manuel Espinosa
Original Articles

Abstract

Deletion of a region of the promiscuous plasmid pLS1 encompassing the initiation signals for the synthesis of the plasmid lagging strand led to plasmid instability in Streptococcus pneumoniae and Bacillus subtilis. This defect could not be alleviated by increasing the number of copies (measured as double-stranded plasmid DNA) to levels similar to those of the wild-type plasmid pLS1. Our results indicate that in the vicinity of, or associated with the single-stranded origin region of pLS1 there is a plasmid component involved in its stable inheritance. Homology was found between the DNA gyrase binding site within the par region of plasmid pSC101 and the pLS1 specific recombination site RSR.

Key words

Minus origin Single-strand origin Plasmid stability Rolling circle replication Single-stranded DNA 

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References

  1. Ballester S, López P, Espinosa M, Alonso JC, Lacks SA (1989) Plasmid structural instability associated with pC194 replication functions. J Bacteriol 171:2271–2277Google Scholar
  2. Ballester S, Alonso JC, López P, Espinosa M (1990) Comparative expression of the pC194 cat gene in Streptococcus pneumoniae, Bacillus subtilis and Escherichia coli. Gene 86:71–79Google Scholar
  3. Biek DP, Cohen SN (1992) Propagation of pSC101 plasmids defective in binding of integration host factor. J Bacteriol 174:785–792Google Scholar
  4. Boe L, Gros MF, te Riele H, Ehrlich SD, Gruss AD (1989) Replication origins of single-stranded DNA plasmid pUB110. J Bacteriol 171:3366–3372Google Scholar
  5. Bron S, Luxen E, Swart P (1988) Instability of recombinant pUB110 plasmids in Bacillus subtilis: plasmid-encoded stability function and effects of DNA inserts. Plasmid 19:231–241Google Scholar
  6. Burdett V (1980) Identification of tetracycline resistant R-plasmids in Streptococcus agalactiae (group B). Antimicrob Agents Chemother 18:753–760Google Scholar
  7. Canosi U, Morelli G, Trautner TA (1978) The relationship between molecular structure and transforming efficiency of some Staphylococcus aureus plasmids isolated from Bacillus subtilis. Mol Gen Genet 166:259–267Google Scholar
  8. de la Campa AG, del Solar G, Espinosa M (1990) Initiation of replication of plasmid pLS 1. The initiator protein RepB acts on two distant DNA regions. J Mol Biol 213:247–262Google Scholar
  9. del Solar G, Espinosa M (1992) The copy number of plasmid pLS1 is regulated by two trans-acting plasmid products: the antisense RNA II and the repressor protein, RepA. Mol Microbiol 6:83–94Google Scholar
  10. del Solar G, Diaz R, Espinosa M (1987a) Replication of the streptococcal plasmid pMV158 and derivatives in cell-free extracts of Escherichia coli. Mol Gen Genet 206:428–435Google Scholar
  11. del Solar G, Puyet A, Espinosa M (1987b) Initiation signals for the conversion of single stranded to double stranded DNA forms in the streptococcal plasmid pLS 1. Nucleic Acids Res 15:5561–5580Google Scholar
  12. del Solar G, de la Campa AG, Perez-Martin J, Choli T, Espinosa M (1989) Purification and characterization of RepA, a protein involved in the copy number control of plasmid pLS 1. Nucleic Acids Res 17:2405–2420Google Scholar
  13. Espinosa M, López P, Perez-Urefia MT, Lacks SA (1982) Interspecific plasmid transfer between Streptococcus pneumoniae and Bacillus subtilis. Mol Gen Genet 188:195–201Google Scholar
  14. Gennaro ML, Novick RP (1986) cmp, a cis-acting plasmid locus that increases interaction between replication origin and initiator protein. J Bacteriol 168:160–166Google Scholar
  15. Gennaro ML, Novick RP (1988) An enhancer of DNA replication. J Bacteriol 170:5709–5717Google Scholar
  16. Gruss AD, Ehrlich SD (1989) The family of highly interrelated single-stranded deoxyribonucleic acid plasmids. Microbiol Rev 53:231–241Google Scholar
  17. Gruss AD, Ross HF, Novick RP (1987) Functional analysis of a palindromic sequence required for normal replication of several, staphylococcal plasmids. Proc Natl Acad Sci USA 84:2165–2169Google Scholar
  18. Khan SA, Novick RP (1983) Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus. Plasmid 10:251–269Google Scholar
  19. Koepsel RR, Murray RW, Khan SA (1986) Sequence-specific interaction between the replication initiator protein of plasmid pT181 and its origin of replication. Proc Natl Acad Sci USA 83:5484–5488Google Scholar
  20. Lacks SA, López P, Greenberg B, Espinosa M (1986) Identification and analysis of genes for tetracycline resistance and replication functions in the broad-host-range plasmid pLSI. J Mol Biol 192:753–765Google Scholar
  21. Leenhouts KJ, Tolner B, Bron S, Kok J, Venema G, Seegers JFML (1991) Nucleotide sequence and characterization of the broadhost-range lactococcal plasmid pWVOI. Plasmid 26:55–66Google Scholar
  22. McKenzie T, Hoshino T, Tanaka T, Sueoka N (1986) The nucleotide sequence of pUB110: some salient features in relation to replication and its regulation. Plasmid 15:93–103Google Scholar
  23. Manen D, Goebel T, Caro L (1990) The par region of pSC101 affects plasmid copy number as well as stability. Mol Microbiol 4:1839–1846Google Scholar
  24. Miller CA, Tucker WT, Meacock PA, Gustafsson P, Cohen SN (1983) Nucleotide sequence of the partition locus of Escherichia coli plasmid pSC101. Gene 24:309–315Google Scholar
  25. Nordström K, Austin SJ (1989) Mechanisms that contribute to the stable segregation of plasmids. Annu Rev Genet 23:37–69Google Scholar
  26. Novick RP (1989) Staphylococcal plasmids and their replication. Anna Rev Microbiol 43:537–565Google Scholar
  27. Novick RP, Projan SJ, Rosenblum W, Edelman I (1984) Staphylococcal plasmid cointegrates are formed by host- and phagemediated general rec systems that act on short regions of homology. Mol Gen Genet 195:374–377Google Scholar
  28. Pérez-Martin J, del Solar G, de la Campa AG, Espinosa M (1988) Three regions in the DNA of plasmid pLSI show sequencedirected static bending. Nucleic Acids Res 16:9113–9126Google Scholar
  29. Priebe SD, Lacks SA (1989) Region of the streptococcal plasmid pMV158 required for conjugative mobilization. J Bacteriol 171:4778–4784Google Scholar
  30. Puyet A, del Solar G, Espinosa M (1988) Identification of the origin and direction of replication of the broad-host-range plasmid pLSI. Nucl Acids Res 16:115–133Google Scholar
  31. Stassi DL, López P, Espinosa M, Lacks SA (1981) Cloning of chromosomal genes in Streptococcus pneumoniae. Proc Natl Acad Sci USA 78:7028–7032Google Scholar
  32. Summers DK (1991) The kinetics of plasmid loss. Trends Biotechnol 9:273–278Google Scholar
  33. te Riele H, Michel B, Ehrlich SD (1986) Are single-stranded circle intermediates in plasmid DNA replication? EMBO 15:631–637Google Scholar
  34. van der Lelie D, Bron S, Venema G, Oskam L (1989) Similarity of minus origins of replication and flanking open reading frames of plasmids pUB110, pTB913 and pMV158. Nucleic Acids Res 17:7283–7294Google Scholar
  35. Wahle E, Kornberg A (1988) The partition locus of plasmid pSCl.01 is a specific binding site for DNA gyrase. EMBO J 7:1889–1895Google Scholar
  36. Xu F, Pearce LE, Yu PL (1991) Genetic analysis of a lactococcal plasmid replicon. Mol Gen Genet 227:33–39Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Gloria del Solar
    • 1
  • Gabriela Kramer
    • 1
  • Sara Ballester
    • 1
  • Manuel Espinosa
    • 1
  1. 1.Centro de Investigaciones BiológicasCSICVelázquez 144MadridSpain

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