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.
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References
Ballester S, López P, Espinosa M, Alonso JC, Lacks SA (1989) Plasmid structural instability associated with pC194 replication functions. J Bacteriol 171:2271–2277
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 86:71–79
Biek DP, Cohen SN (1992) Propagation of pSC101 plasmids defective in binding of integration host factor. J Bacteriol 174:785–792
Boe L, Gros MF, te Riele H, Ehrlich SD, Gruss AD (1989) Replication origins of single-stranded DNA plasmid pUB110. J Bacteriol 171:3366–3372
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 19:231–241
Burdett V (1980) Identification of tetracycline resistant R-plasmids in Streptococcus agalactiae (group B). Antimicrob Agents Chemother 18:753–760
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 166:259–267
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 213:247–262
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 6:83–94
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 206:428–435
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 15:5561–5580
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 17:2405–2420
Espinosa M, López P, Perez-Urefia MT, Lacks SA (1982) Interspecific plasmid transfer between Streptococcus pneumoniae and Bacillus subtilis. Mol Gen Genet 188:195–201
Gennaro ML, Novick RP (1986) cmp, a cis-acting plasmid locus that increases interaction between replication origin and initiator protein. J Bacteriol 168:160–166
Gennaro ML, Novick RP (1988) An enhancer of DNA replication. J Bacteriol 170:5709–5717
Gruss AD, Ehrlich SD (1989) The family of highly interrelated single-stranded deoxyribonucleic acid plasmids. Microbiol Rev 53:231–241
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 84:2165–2169
Khan SA, Novick RP (1983) Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus. Plasmid 10:251–269
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 83:5484–5488
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 192:753–765
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 26:55–66
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 15:93–103
Manen D, Goebel T, Caro L (1990) The par region of pSC101 affects plasmid copy number as well as stability. Mol Microbiol 4:1839–1846
Miller CA, Tucker WT, Meacock PA, Gustafsson P, Cohen SN (1983) Nucleotide sequence of the partition locus of Escherichia coli plasmid pSC101. Gene 24:309–315
Nordström K, Austin SJ (1989) Mechanisms that contribute to the stable segregation of plasmids. Annu Rev Genet 23:37–69
Novick RP (1989) Staphylococcal plasmids and their replication. Anna Rev Microbiol 43:537–565
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 195:374–377
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 16:9113–9126
Priebe SD, Lacks SA (1989) Region of the streptococcal plasmid pMV158 required for conjugative mobilization. J Bacteriol 171:4778–4784
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 16:115–133
Stassi DL, López P, Espinosa M, Lacks SA (1981) Cloning of chromosomal genes in Streptococcus pneumoniae. Proc Natl Acad Sci USA 78:7028–7032
Summers DK (1991) The kinetics of plasmid loss. Trends Biotechnol 9:273–278
te Riele H, Michel B, Ehrlich SD (1986) Are single-stranded circle intermediates in plasmid DNA replication? EMBO 15:631–637
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 17:7283–7294
Wahle E, Kornberg A (1988) The partition locus of plasmid pSCl.01 is a specific binding site for DNA gyrase. EMBO J 7:1889–1895
Xu F, Pearce LE, Yu PL (1991) Genetic analysis of a lactococcal plasmid replicon. Mol Gen Genet 227:33–39
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Communicated by R. Devoret
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del Solar, G., Kramer, G., Ballester, S. et al. Replication of the promiscuous plasmid pLSI: a region encompassing the minus origin of replication is associated with stable plasmid inheritance. Molec. Gen. Genet. 241, 97–105 (1993). https://doi.org/10.1007/BF00280206
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DOI: https://doi.org/10.1007/BF00280206