The replicon of pSW800 from

The GenBank accession number for the sequence of the minimal replicon of pSW800 is AF310258.

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Abstract

A 2019 bp DNA fragment containing the replicon of pSW800 from SW2 was cloned and characterized. This replicon contains two genes – and , which encode a 365 kDa replication initiation protein (RepA) and a peptide of 18 aa, respectively. These two genes overlap by 8 bases with situated upstream. The replicon also transcribes an antisense RNA (RNAI) that inhibits the expression of and . The ribosome-binding sequence (RBS) of is likely to be hidden in a stem–loop structure, inhibiting the translation of . Furthermore, translation of is likely to disrupt the stem–loop structure, which is one of the criteria allowing the translation of to begin. A mutagenesis study revealed that a sequence (5′-GCACGGG-3′) located 111 nt upstream from is crucial; mutation of this sequence prevented the translation of . Additionally, this region and the stem–loop structure containing the RBS of may form an RNA pseudoknot. Results in this study demonstrate that a mechanism similar to that regulating plasmid replication in the IncB, IncIα and IncL/M groups also regulates pSW800 replication.

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2001-10-01
2024-03-28
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References

  1. Aiba H., Adhya S., de Crombrugghe B. 1981; Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
    [Google Scholar]
  2. Asano K., Mizobuchi K. 1998a; Copy number control of Incα plasmid ColIb-P9 by competition between pseudoknot formation and antisense RNA binding at a specific RNA site. EMBO J 17:5201–5213 [CrossRef]
    [Google Scholar]
  3. Asano K., Mizobuchi K. 1998b; An RNA pseudoknot as the molecular switch for translation of the repZ gene encoding the replication initiator of IncIα plasmid ColIb-P9. J Biol Chem 273:11815–11825 [CrossRef]
    [Google Scholar]
  4. Asano K., Mizobuchi K. 2000; Structural analysis of late intermediate complex formed between plasmid ColIb-P9 Inc RNA and its target RNA. How does a single antisense RNA repress translation of two genes at different rates?. J Biol Chem 275:1269–1274 [CrossRef]
    [Google Scholar]
  5. Asano K., Niimi T., Yokoyama S., Mizobuchi K. 1998; Structural basis for binding of the plasmid ColIb-P9 antisense Inc RNA to its target RNA with the 5′-rUUGGCG-3′ motif in the loop sequence. J Biol Chem 273:11826–11838 [CrossRef]
    [Google Scholar]
  6. Asano K., Hama C., Inoue S., Moriwaki H., Mizobuchi K. 1999; The plasmid ColIb-P9 antisense Inc RNA controls expression of the RepZ replication protein and its positive regulator repY with different mechanisms. J Biol Chem 274:17924–17933 [CrossRef]
    [Google Scholar]
  7. Athanasopoulos V., Praszkier J., Pittard A. J. 1995; The replication of an IncL/M plasmid is subject to antisense control. J Bacteriol 177:4730–4741
    [Google Scholar]
  8. Athanasopoulos V., Praszkier J., Pittard A. J. 1999; Analysis of elements involved in pseudoknot-dependent expression and regulation of the repA gene of an IncL/M plasmid. J Bacteriol 181:1811–1819
    [Google Scholar]
  9. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523 [CrossRef]
    [Google Scholar]
  10. Blomberg P., Wagner E. G. H., Nördstrom K. 1992; Replication control of plasmid R1: RepA synthesis is regulated by CopA RNA through inhibition of leader peptide translation. EMBO J 11:2675–2683
    [Google Scholar]
  11. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113 [CrossRef]
    [Google Scholar]
  12. Boyer H. W., Roulland-Dessoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli . J Mol Biol 41:459–472 [CrossRef]
    [Google Scholar]
  13. Burian J., Stuchlik S., Kay W. W. 1999; Replication control of a small cryptic plasmid of Escherichia coli . J Mol Biol 294:49–65 [CrossRef]
    [Google Scholar]
  14. Chang P. J., Chang Y. S., Liu S. T. 1998; Role of Rta in the translation of bicistronic BZLF1 of Epstein–Barr virus. J Virol 72:5128–5136
    [Google Scholar]
  15. Coplin D. L., Rowan R. G., Chisholm D. A., Whitmoyer R. E. 1981; Characterization of plasmids in Erwinia stewartii . Appl Environ Microbiol 42:599–604
    [Google Scholar]
  16. Coplin D. L., Frederick R. D., McCammon S. L. 1985; Characterization of a conjugative plasmid from Erwinia stewartii . J Gen Microbiol 131:2985–2991
    [Google Scholar]
  17. Franch T., Thisted T., Gerdes K. 1999; Ribonuclease III processing of coaxially stacked RNA helices. J Biol Chem 274:26572–26578 [CrossRef]
    [Google Scholar]
  18. Frederick R. D., Coplin D. L. 1986; Transformation of Escherichia coli by plasmid DNA from Erwinia stewartii . Mol Plant Pathol 76:1353–1356
    [Google Scholar]
  19. Fu J. F., Chang H. C., Chen Y. M., Chang Y. S., Liu S. T. 1995; Sequence analysis of an Erwinia stewartii plasmid, pSW100. Plasmid 34:75–84 [CrossRef]
    [Google Scholar]
  20. Fu J. F., Chang H. C., Chen Y. M., Chang Y. S., Liu S. T. 1996; Characterization of the replicon of plasmid pSW500 of Erwinia stewartii . Mol Gen Genet 250:699–704
    [Google Scholar]
  21. Fu J. F., Ying S. W., Liu S. T. 1997; Cloning and characterization of the ori region of pSW1200 of Erwinia stewartii : similarity with plasmid P1. Plasmid 38:141–147 [CrossRef]
    [Google Scholar]
  22. Fu J. F., Hu J. M., Chang Y. S., Liu S. T. 1998; Isolation and characterization of plasmid pSW200 from Erwinia stewartii . Plasmid 40:100–112 [CrossRef]
    [Google Scholar]
  23. Hama C., Takizawa T., Moriwaki H., Urasaki Y., Mizobuchi K. 1990; Organization of the replication control region of plasmid ColIb-P9. J Bacteriol 172:1983–1991
    [Google Scholar]
  24. Hamilton C. M., Aldea M., Washburn B. K., Babitzke P., Kushner S. R. 1989; New method for generating deletions and gene replacements in Escherichia coli . J Bacteriol 171:4617–4622
    [Google Scholar]
  25. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59 [CrossRef]
    [Google Scholar]
  26. Kado C. I., Liu S. T. 1981; Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145:1365–1373
    [Google Scholar]
  27. Lin T. P., Chen C. L., Chang L. K., Tschen J. S., Liu S. T. 1999; Functional and transcriptional analyses of a fengycin synthetase gene, fenC , from Bacillus subtilis . J Bacteriol 181:5060–5067
    [Google Scholar]
  28. Masui Y., Coleman J., Inouye M. 1983 Multipurpose Expression Cloning Vehicles in Escherichia coli New York: Academic Press;
    [Google Scholar]
  29. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Oka A., Sugisaki H., Takanami M. 1981; Nucleotide sequence of the kanamycin resistance transposon Tn 903 . J Mol Biol 147:217–226 [CrossRef]
    [Google Scholar]
  31. Praszkier J., Wilson I. W., Pittard A. J. 1992; Mutations affecting translational coupling between the rep genes of an IncB miniplasmid. J Bacteriol 174:2376–2383
    [Google Scholar]
  32. Saadi S., Maas W. K., Hill D. F., Bergquist P. L. 1987; Nucleotide sequence analysis of RepFIC, a basic replicon present in IncFI plasmids P307 and F, and its relation to the RepA replicon of IncFII plasmids. J Bacteriol 169:1836–1846
    [Google Scholar]
  33. Sambrook J., Russell D. 2000 Molecular Cloning: a Laboratory Manual , 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467 [CrossRef]
    [Google Scholar]
  35. Siemering K. R., Praszkier J., Pittard A. J. 1993; Interaction between the antisense and target RNAs involved in the regulation of IncB plasmid replication. J Bacteriol 175:2895–2906
    [Google Scholar]
  36. del Solar G., Giraldo R., Ruiz-Echevarria M. J., Espinosa M., Diaz-Orejas R. 1998; Replication and control of circular bacterial plasmids. Microbiol Mol Biol Rev 62:434–464
    [Google Scholar]
  37. Wu R. P., Womble D. D., Rownd R. H. 1985; Incompatibility mutants of IncFII plasmid NR1 and their effect on replication control. J Bacteriol 163:973–982
    [Google Scholar]
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