Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Nov;164(2):556–562. doi: 10.1128/jb.164.2.556-562.1985

Characterization of the in vivo RNA product of the pOUT promoter of IS10R.

Y Lee, F J Schmidt
PMCID: PMC214288  PMID: 2414269

Abstract

We characterized a single RNA species (RNAout1) which was the major in vivo RNA made from pOUT of IS10R. RNAout1 was 70 nucleotides long; its 5' end corresponded exactly to the in vitro start of pOUT transcription. The concentration of RNAout1 was estimated at 5 to 10 molecules per cell containing the single-copy plasmid NR1. RNA sequences from pOUT of IS10L were detected at a much lower (less than one molecule per cell) steady-state concentration and may be preferentially degraded in vivo. We suggest that the low level of the IS10L transcript led to the inability of IS10L sequences to translationally inhibit Tn10 transposition.

Full text

PDF
559

Images in this article

558Image
on p.558
559Image
on p.559
560Image
on p.560
561Image
on p.561

Selected References

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

  1. Apirion D., Neil J., Ko T. S., Watson N. Metabolism of ribosomal RNA in mutants of Escherichia coli doubly defective in ribonuclease III and the transcription termination factor rho. Genetics. 1978 Sep;90(1):19–35. doi: 10.1093/genetics/90.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ciampi M. S., Schmid M. B., Roth J. R. Transposon Tn10 provides a promoter for transcription of adjacent sequences. Proc Natl Acad Sci U S A. 1982 Aug;79(16):5016–5020. doi: 10.1073/pnas.79.16.5016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Wilde M., Davies J. E., Schmidt F. J. Low molecular weight RNA species encoded by a multiple drug resistance plasmid. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3673–3677. doi: 10.1073/pnas.75.8.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ebina Y., Nakazawa A. Cyclic AMP-dependent initiation and rho-dependent termination of colicin E1 gene transcription. J Biol Chem. 1983 Jun 10;258(11):7072–7078. [PubMed] [Google Scholar]
  6. Halling S. M., Simons R. W., Way J. C., Walsh R. B., Kleckner N. DNA sequence organization of IS10-right of Tn10 and comparison with IS10-left. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2608–2612. doi: 10.1073/pnas.79.8.2608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hillen W., Klock G., Kaffenberger I., Wray L. V., Reznikoff W. S. Purification of the TET repressor and TET operator from the transposon Tn10 and characterization of their interaction. J Biol Chem. 1982 Jun 10;257(11):6605–6613. [PubMed] [Google Scholar]
  8. Johnson R. A., Walseth T. F. The enzymatic preparation of [alpha-32P]ATP, [alpha-32P]GTP, [32P]cAMP, and [32P]cGMP, and their use in the assay of adenylate and guanylate cyclases and cyclic nucleotide phosphodiesterases. Adv Cyclic Nucleotide Res. 1979;10:135–167. [PubMed] [Google Scholar]
  9. Jorgensen R. A., Reznikoff W. S. Organization of structural and regulatory genes that mediate tetracycline resistance in transposon Tn10. J Bacteriol. 1979 Jun;138(3):705–714. doi: 10.1128/jb.138.3.705-714.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kleckner N., Chan R. K., Tye B. K., Botstein D. Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol. 1975 Oct 5;97(4):561–575. doi: 10.1016/s0022-2836(75)80059-3. [DOI] [PubMed] [Google Scholar]
  11. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Motamedi H., Lee K., Nichols L., Schmidt F. J. An RNA species involved in Escherichia coli ribonuclease P activity. Gene cloning and effect on transfer RnA synthesis in vivo. J Mol Biol. 1982 Dec 15;162(3):535–550. doi: 10.1016/0022-2836(82)90387-4. [DOI] [PubMed] [Google Scholar]
  15. Nishimura S. Minor components in transfer RNA: their characterization, location, and function. Prog Nucleic Acid Res Mol Biol. 1972;12:49–85. [PubMed] [Google Scholar]
  16. Schmidt F. J., Jorgensen R. A., de Wilde M., Davies J. E. A specific tetracycline-induced, low-molecular-weight RNA encoded by the inverted repeat of Tn10 (IS10). Plasmid. 1981 Jul;6(1):148–150. doi: 10.1016/0147-619x(81)90061-5. [DOI] [PubMed] [Google Scholar]
  17. Simons R. W., Hoopes B. C., McClure W. R., Kleckner N. Three promoters near the termini of IS10: pIN, pOUT, and pIII. Cell. 1983 Sep;34(2):673–682. doi: 10.1016/0092-8674(83)90400-2. [DOI] [PubMed] [Google Scholar]
  18. Simons R. W., Kleckner N. Translational control of IS10 transposition. Cell. 1983 Sep;34(2):683–691. doi: 10.1016/0092-8674(83)90401-4. [DOI] [PubMed] [Google Scholar]
  19. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  20. Wray L. V., Jr, Reznikoff W. S. Identification of repressor binding sites controlling expression of tetracycline resistance encoded by Tn10. J Bacteriol. 1983 Dec;156(3):1188–1191. doi: 10.1128/jb.156.3.1188-1191.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES