Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1982;1(3):323–328. doi: 10.1002/j.1460-2075.1982.tb01168.x

Convergent transcription interferes with expression of the copy number control gene, copA, from plasmid R1

Peter Stougaard 1,1, Janice Light 1,2, Søren Molin 1,*
PMCID: PMC553043  PMID: 16453418

Abstract

The copy number control gene, copA, of plasmid R1 codes for an 80-nucleotide untranslatable RNA. In Escherichia coli minicells, some copA hybrid plasmids and R1 miniplasmids also express a transcript of ˜200 nucleotides. Only the small RNA mediates the CopA phenotype. The switch between 80- and 200-nucleotide RNA synthesis is shown to be caused by convergent transcription; if transcription proceeds in both directions the inactive larger RNA is synthesised; the active small RNA is formed when copA transcription is not opposed by transcription from the other direction. The data presented indicate that convergent transcription interferes with copA expression by abolishing or reducing normal copA transcription termination.

Keywords: E. coli minicells, oppositely directed transcription, plasmid incompatibility, transcription termination

Full text

PDF
323

Images in this article

Selected References

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

  1. An G., Friesen J. D. Plasmid vehicles for direct cloning of Escherichia coli promoters. J Bacteriol. 1979 Nov;140(2):400–407. doi: 10.1128/jb.140.2.400-407.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Backman K., Ptashne M. Maximizing gene expression on a plasmid using recombination in vitro. Cell. 1978 Jan;13(1):65–71. doi: 10.1016/0092-8674(78)90138-1. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  5. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Danbara H., Brady G., Timmis J. K., Timmis K. N. Regulation of DNA replication: "target" determinant of the replication control elements of plasmid R6-5 lies within a control element gene. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4699–4703. doi: 10.1073/pnas.78.8.4699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Danbara H., Timmis J. K., Lurz R., Timmis K. N. Plasmid replication functions: two distinct segments of plasmid R1, RepA and RepD, express incompatibility and are capable of autonomous replication. J Bacteriol. 1980 Dec;144(3):1126–1138. doi: 10.1128/jb.144.3.1126-1138.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  10. Heidecker G., Messing J., Gronenborn B. A versatile primer for DNA sequencing in the M13mp2 cloning system. Gene. 1980 Jun;10(1):69–73. doi: 10.1016/0378-1119(80)90145-6. [DOI] [PubMed] [Google Scholar]
  11. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  12. Kennedy N., Beutin L., Achtman M., Skurray R., Rahmsdorf U., Herrlich P. Conjugation proteins encoded by the F sex factor. Nature. 1977 Dec 15;270(5638):580–585. doi: 10.1038/270580a0. [DOI] [PubMed] [Google Scholar]
  13. Light J., Molin S. Replication control functions of plasmid R1 act as inhibitors of expression of a gene required for replication. Mol Gen Genet. 1981;184(1):56–61. doi: 10.1007/BF00271195. [DOI] [PubMed] [Google Scholar]
  14. Meynell E., Datta N. Mutant drug resistant factors of high transmissibility. Nature. 1967 May 27;214(5091):885–887. doi: 10.1038/214885a0. [DOI] [PubMed] [Google Scholar]
  15. Molin S., Nordström K. Control of plasmid R1 replication: functions involved in replication, copy number control, incompatibility, and switch-off of replication. J Bacteriol. 1980 Jan;141(1):111–120. doi: 10.1128/jb.141.1.111-120.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Molin S., Stougaard P., Light J., Nordström M., Nordström K. Isolation and characterization of new copy mutants of plasmid R1, and identification of a polypeptide involved in copy number control. Mol Gen Genet. 1981;181(1):123–130. doi: 10.1007/BF00339015. [DOI] [PubMed] [Google Scholar]
  17. Molin S., Stougaard P., Uhlin B. E., Gustafsson P., Nordström K. Clustering of genes involved in replication, copy number control, incompatibility, and stable maintenance of the resistance plasmid R1drd-19. J Bacteriol. 1979 Apr;138(1):70–79. doi: 10.1128/jb.138.1.70-79.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rosen J., Ryder T., Inokuchi H., Ohtsubo H., Ohtsubo E. Genes and sites involved in replication and incompatibility of an R100 plasmid derivative based on nucleotide sequence analysis. Mol Gen Genet. 1980;179(3):527–537. doi: 10.1007/BF00271742. [DOI] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Stougaard P., Molin S., Nordström K., Hansen F. G. The nucleotide sequence of the replication control region of the resistance plasmid R1drd-19. Mol Gen Genet. 1981;181(1):116–122. doi: 10.1007/BF00339014. [DOI] [PubMed] [Google Scholar]
  21. Stougaard P., Molin S., Nordström K. RNAs involved in copy-number control and incompatibility of plasmid R1. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6008–6012. doi: 10.1073/pnas.78.10.6008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stougaard P., Molin S. Vertical dye-buoyant density gradients for rapid analysis and preparation of plasmid DNA. Anal Biochem. 1981 Nov 15;118(1):191–193. doi: 10.1016/0003-2697(81)90177-9. [DOI] [PubMed] [Google Scholar]
  23. Ward D. F., Murray N. E. Convergent transcription in bacteriophage lambda: interference with gene expression. J Mol Biol. 1979 Sep 15;133(2):249–266. doi: 10.1016/0022-2836(79)90533-3. [DOI] [PubMed] [Google Scholar]
  24. Yoshikawa M. Identification and mapping of the replication genes of an R factor, R100-1, integrated into the chromosome of Escherichia coli K-12. J Bacteriol. 1974 Jun;118(3):1123–1131. doi: 10.1128/jb.118.3.1123-1131.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES