Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Sep 12;16(17):8411–8431. doi: 10.1093/nar/16.17.8411

Recognition of prokaryotic transcription terminators by spinach chloroplast RNA polymerase.

L J Chen 1, E M Orozco Jr 1
PMCID: PMC338567  PMID: 2843817

Abstract

To determine whether chloroplast RNA polymerase will accurately terminate transcription in vitro, we have fused the spinach chloroplast rbcL promoter to the 3' end of the rbcL gene as well as to various factor independent transcription terminators from E. coli. Transcription of the rbcL minigene did not result in production of the expected 265 nucleotide RNA. However, the spinach chloroplast RNA polymerase did terminate transcription with varying efficiency at the thra, rrnB, rrnC and gene 32 terminators. The most efficient transcription termination was observed for the threonine attenuator. For each of the prokaryotic terminators, the chloroplast enzyme ceased transcription at essentially the same position as the E. coli RNA polymerase. These data indicate that the transcription termination process in chloroplasts has some features in common with the mechanism used in prokaryotes.

Full text

PDF
8411

Images in this article

8418Image
on p.8418
8421Image
on p.8421
8422Image
on p.8422
8424Image
on p.8424

Selected References

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

  1. Brosius J., Dull T. J., Sleeter D. D., Noller H. F. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol. 1981 May 15;148(2):107–127. doi: 10.1016/0022-2836(81)90508-8. [DOI] [PubMed] [Google Scholar]
  2. Chapman K. A., Burgess R. R. Construction of bacteriophage T7 late promoters with point mutations and characterization by in vitro transcription properties. Nucleic Acids Res. 1987 Jul 10;15(13):5413–5432. doi: 10.1093/nar/15.13.5413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Freier S. M., Kierzek R., Jaeger J. A., Sugimoto N., Caruthers M. H., Neilson T., Turner D. H. Improved free-energy parameters for predictions of RNA duplex stability. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9373–9377. doi: 10.1073/pnas.83.24.9373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gardner J. F. Initiation, pausing, and termination of transcription in the threonine operon regulatory region of Escherichia coli. J Biol Chem. 1982 Apr 10;257(7):3896–3904. [PubMed] [Google Scholar]
  5. Gruissem W., Zurawski G. Analysis of promoter regions for the spinach chloroplast rbcL, atpB and psbA genes. EMBO J. 1985 Dec 16;4(13A):3375–3383. doi: 10.1002/j.1460-2075.1985.tb04093.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gruissem W., Zurawski G. Identification and mutational analysis of the promoter for a spinach chloroplast transfer RNA gene. EMBO J. 1985 Jul;4(7):1637–1644. doi: 10.1002/j.1460-2075.1985.tb03831.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hanley-Bowdoin L., Orozco E. M., Jr, Chua N. H. In vitro synthesis and processing of a maize chloroplast transcript encoded by the ribulose 1,5-bisphosphate carboxylase large subunit gene. Mol Cell Biol. 1985 Oct;5(10):2733–2745. doi: 10.1128/mcb.5.10.2733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Holschuh K., Bottomley W., Whitfeld P. R. Structure of the spinach chloroplast genes for the D2 and 44 kd reaction-centre proteins of photosystem II and for tRNASer (UGA). Nucleic Acids Res. 1984 Dec 11;12(23):8819–8834. doi: 10.1093/nar/12.23.8819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Krisch H. M., Allet B. Nucleotide sequences involved in bacteriophage T4 gene 32 translational self-regulation. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4937–4941. doi: 10.1073/pnas.79.16.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lynn S. P., Gardner J. F., Reznikoff W. S. Attenuation regulation in the thr operon of Escherichia coli K-12: molecular cloning and transcription of the controlling region. J Bacteriol. 1982 Oct;152(1):363–371. doi: 10.1128/jb.152.1.363-371.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mead D. A., Szczesna-Skorupa E., Kemper B. Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1986 Oct-Nov;1(1):67–74. doi: 10.1093/protein/1.1.67. [DOI] [PubMed] [Google Scholar]
  12. Mott J. E., Galloway J. L., Platt T. Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination. EMBO J. 1985 Jul;4(7):1887–1891. doi: 10.1002/j.1460-2075.1985.tb03865.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Orozco E. M., Jr, Mullet J. E., Chua N. H. An in vitro system for accurate transcription initiation of chloroplast protein genes. Nucleic Acids Res. 1985 Feb 25;13(4):1283–1302. doi: 10.1093/nar/13.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Orozco E. M., Jr, Mullet J. E., Hanley-Bowdoin L., Chua N. H. In vitro transcription of chloroplast protein genes. Methods Enzymol. 1986;118:232–253. doi: 10.1016/0076-6879(86)18076-1. [DOI] [PubMed] [Google Scholar]
  15. Platt T. Transcription termination and the regulation of gene expression. Annu Rev Biochem. 1986;55:339–372. doi: 10.1146/annurev.bi.55.070186.002011. [DOI] [PubMed] [Google Scholar]
  16. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  17. Schmidt M. C., Chamberlin M. J. nusA protein of Escherichia coli is an efficient transcription termination factor for certain terminator sites. J Mol Biol. 1987 Jun 20;195(4):809–818. doi: 10.1016/0022-2836(87)90486-4. [DOI] [PubMed] [Google Scholar]
  18. Shinozaki K., Ohme M., Tanaka M., Wakasugi T., Hayashida N., Matsubayashi T., Zaita N., Chunwongse J., Obokata J., Yamaguchi-Shinozaki K. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 1986 Sep;5(9):2043–2049. doi: 10.1002/j.1460-2075.1986.tb04464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sijben-Müller G., Hallick R. B., Alt J., Westhoff P., Herrmann R. G. Spinach plastid genes coding for initiation factor IF-1, ribosomal protein S11 and RNA polymerase alpha-subunit. Nucleic Acids Res. 1986 Jan 24;14(2):1029–1044. doi: 10.1093/nar/14.2.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stern D. B., Gruissem W. Control of plastid gene expression: 3' inverted repeats act as mRNA processing and stabilizing elements, but do not terminate transcription. Cell. 1987 Dec 24;51(6):1145–1157. doi: 10.1016/0092-8674(87)90600-3. [DOI] [PubMed] [Google Scholar]
  21. Young R. A. Transcription termination in the Escherichia coli ribosomal RNA operon rrnC. J Biol Chem. 1979 Dec 25;254(24):12725–12731. [PubMed] [Google Scholar]
  22. Zurawski G., Bohnert H. J., Whitfeld P. R., Bottomley W. Nucleotide sequence of the gene for the M(r) 32,000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of M(r) 38,950. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7699–7703. doi: 10.1073/pnas.79.24.7699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zurawski G., Bottomley W., Whitfeld P. R. Structures of the genes for the beta and epsilon subunits of spinach chloroplast ATPase indicate a dicistronic mRNA and an overlapping translation stop/start signal. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6260–6264. doi: 10.1073/pnas.79.20.6260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zurawski G., Perrot B., Bottomley W., Whitfeld P. R. The structure of the gene for the large subunit of ribulose 1,5-bisphosphate carboxylase from spinach chloroplast DNA. Nucleic Acids Res. 1981 Jul 24;9(14):3251–3270. doi: 10.1093/nar/9.14.3251. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES