Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Oct;79(20):6313–6317. doi: 10.1073/pnas.79.20.6313

Control of ColE1 DNA replication: the rop gene product negatively affects transcription from the replication primer promoter.

G Cesareni, M A Muesing, B Polisky
PMCID: PMC347111  PMID: 6183660

Abstract

A 600-base-pair region essential for ColE1 and pMBl plasmid replication contains two promoters responsible for the synthesis of two RNA molecules central to copy number control. One promoter directs synthesis of the primer RNA precursor. The second promoter directs the synthesis of a small RNA molecule, RNAl, which acts in trans to inhibit processing of the RNA primer precursor. We have fused each promoter to the beta-galactosidase structural gene contained in a lambda phage. Expression of the RNAl promoter in lysogens is not influenced by the presence of wild-type pMBl or ColEl plasmids residing in the cell. Transcription from the RNA primer promoter, however, is repressed by the product of a trans-acting plasmid gene product, which we have designated rop (for repressor of primer). The rop gene maps downstream from the replication origin in a region that encodes a polypeptide of 63 amino acids whose sequence is completely conserved in pMBl and ColE1. We propose that this polypeptide is the rop gene product and that it regulates plasmid DNA replication by modulating the initiation of transcription of the primer RNA precursor.

Full text

PDF
6315

Images in this article

6316Image
on p.6316

Selected References

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

  1. Armstrong K. A., Hershfield V., Helinski D. R. Gene cloning and containment properties of plasmid Col E1 and its derivatives. Science. 1977 Apr 8;196(4286):172–174. doi: 10.1126/science.322277. [DOI] [PubMed] [Google Scholar]
  2. Backman K., Betlach M., Boyer H. W., Yanofsky S. Genetic and physical studies on the replication of ColE1-type plasmids. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):69–76. doi: 10.1101/sqb.1979.043.01.012. [DOI] [PubMed] [Google Scholar]
  3. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  4. Betlach M., Hershfield V., Chow L., Brown W., Goodman H., Boyer H. W. A restriction endonuclease analysis of the bacterial plasmid controlling the ecoRI restriction and modification of DNA. Fed Proc. 1976 Jul;35(9):2037–2043. [PubMed] [Google Scholar]
  5. Brenner S., Cesareni G., Karn J. Phasmids: hybrids between ColE1 plasmids and E. coli bacteriophage lambda. Gene. 1982 Jan;17(1):27–44. doi: 10.1016/0378-1119(82)90098-1. [DOI] [PubMed] [Google Scholar]
  6. Cesareni G. The target of the negative regulator of pMB1 replication overlaps with part of the repressor coding sequence. Mol Gen Genet. 1981;184(1):40–45. doi: 10.1007/BF00271192. [DOI] [PubMed] [Google Scholar]
  7. Conrad S. E., Campbell J. L. Role of plasmid-coded RNA and ribonuclease III in plasmid DNA replication. Cell. 1979 Sep;18(1):61–71. doi: 10.1016/0092-8674(79)90354-4. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Lacatena R. M., Cesareni G. Base pairing of RNA I with its complementary sequence in the primer precursor inhibits ColE1 replication. Nature. 1981 Dec 17;294(5842):623–626. doi: 10.1038/294623a0. [DOI] [PubMed] [Google Scholar]
  10. Maurer R., Meyer B., Ptashne M. Gene regulation at the right operator (OR) bacteriophage lambda. I. OR3 and autogenous negative control by repressor. J Mol Biol. 1980 May 15;139(2):147–161. doi: 10.1016/0022-2836(80)90302-2. [DOI] [PubMed] [Google Scholar]
  11. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  12. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Muesing M., Tamm J., Shepard H. M., Polisky B. A single base-pair alteration is responsible for the DNA overproduction phenotype of a plasmid copy-number mutant. Cell. 1981 Apr;24(1):235–242. doi: 10.1016/0092-8674(81)90519-5. [DOI] [PubMed] [Google Scholar]
  14. Oka A., Nomura N., Morita M., Sugisaki H., Sugimoto K., Takanami M. Nucleotide sequence of small ColE1 derivatives: structure of the regions essential for autonomous replication and colicin E1 immunity. Mol Gen Genet. 1979 May 4;172(2):151–159. doi: 10.1007/BF00268276. [DOI] [PubMed] [Google Scholar]
  15. Reeve J. N. Mucopeptide biosynthesis by minicells of Escherichia coli. J Bacteriol. 1977 Jul;131(1):363–365. doi: 10.1128/jb.131.1.363-365.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. So M., Gill R., Falkow S. The generation of a ColE1-Apr cloning vehicle which allows detection of inserted DNA. Mol Gen Genet. 1975 Dec 30;142(3):239–249. doi: 10.1007/BF00425649. [DOI] [PubMed] [Google Scholar]
  18. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  19. Tomizawa J., Itoh T. Plasmid ColE1 incompatibility determined by interaction of RNA I with primer transcript. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6096–6100. doi: 10.1073/pnas.78.10.6096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tomizawa J., Itoh T., Selzer G., Som T. Inhibition of ColE1 RNA primer formation by a plasmid-specified small RNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1421–1425. doi: 10.1073/pnas.78.3.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Veltkamp E., van de Pol H., Stuitje A. R., van den Elzen P. J., Nijkamp H. J. Replication and gene functions of the bacteriocinogenic plasmid CloDF13. Contrib Microbiol Immunol. 1979;6:111–121. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES