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. 1986 Sep;5(9):2043–2049. doi: 10.1002/j.1460-2075.1986.tb04464.x

The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression

K Shinozaki *, M Ohme *, M Tanaka *, T Wakasugi *, N Hayashida *, T Matsubayashi *, N Zaita *, J Chunwongse *, J Obokata *, K Yamaguchi-Shinozaki *, C Ohto *, K Torazawa *, B Y Meng *, M Sugita *,1, H Deno *,2, T Kamogashira *,3, K Yamada *,4, J Kusuda 5, F Takaiwa *,6, A Kato *,6, N Tohdoh *,7, H Shimada *,8, M Sugiura *
PMCID: PMC1167080  PMID: 16453699

Abstract

The complete nucleotide sequence (155 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA has been determined. It contains two copies of an identical 25 339 bp inverted repeat, which are separated by a 86 684 bp and a 18 482 bp single-copy region. The genes for 4 different rRNAs, 30 different tRNAs, 39 different proteins and 11 other predicted protein coding genes have been located. Among them, 15 genes contain introns. Blot hybridization revealed that all rRNA and tRNA genes and 27 protein genes so far analysed are transcribed in the chloroplast and that primary transcripts of the split genes hitherto examined are spliced. Five sequences coding for proteins homologous to components of the respiratory-chain NADH dehydrogenase from human mitochondria have been found. The 30 tRNAs predicted from their genes are sufficient to read all codons if the `two out of three' and `U:N wobble' mechanisms operate in the chloroplast. Two sequences which autonomously replicate in yeast have also been mapped. The sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.

Keywords: DNA sequence, gene map, intron, tobacco chloroplast, transcription

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Selected References

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  1. Barrell B. G., Anderson S., Bankier A. T., de Bruijn M. H., Chen E., Coulson A. R., Drouin J., Eperon I. C., Nierlich D. P., Roe B. A. Different pattern of codon recognition by mammalian mitochondrial tRNAs. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3164–3166. doi: 10.1073/pnas.77.6.3164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Capel M. S., Bourque D. P. Characterization of Nicotiana tabacum chloroplast and cytoplasmic ribosomal proteins. J Biol Chem. 1982 Jul 10;257(13):7746–7755. [PubMed] [Google Scholar]
  3. Chomyn A., Mariottini P., Cleeter M. W., Ragan C. I., Matsuno-Yagi A., Hatefi Y., Doolittle R. F., Attardi G. Six unidentified reading frames of human mitochondrial DNA encode components of the respiratory-chain NADH dehydrogenase. Nature. 1985 Apr 18;314(6012):592–597. doi: 10.1038/314592a0. [DOI] [PubMed] [Google Scholar]
  4. Cozens A. L., Walker J. E., Phillips A. L., Huttly A. K., Gray J. C. A sixth subunit of ATP synthase, an F(0) component, is encoded in the pea chloroplast genome. EMBO J. 1986 Feb;5(2):217–222. doi: 10.1002/j.1460-2075.1986.tb04201.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deno H., Kato A., Shinozaki K., Sugiura M. Nucleotide sequences of tobacco chloroplast genes for elongator tRNAMet and tRNAVal (UAC): the tRNAVal (UAC) gene contains a long intron. Nucleic Acids Res. 1982 Dec 11;10(23):7511–7520. doi: 10.1093/nar/10.23.7511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deno H., Shinozaki K., Sugiura M. Nucleotide sequence of tobacco chloroplast gene for the alpha subunit of proton-translocating ATPase. Nucleic Acids Res. 1983 Apr 11;11(7):2185–2191. doi: 10.1093/nar/11.7.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deno H., Sugiura M. Chloroplast tRNA gene contains a long intron in the D stem: Nucleotide sequences of tobacco chloroplast genes for tRNA (UCC) and tRNA (UCU). Proc Natl Acad Sci U S A. 1984 Jan;81(2):405–408. doi: 10.1073/pnas.81.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Deno H., Sugiura M. The nucleotide sequences of tRNASer (GCU) and tRNAGln (UUG) genes from tobacco chloroplasts. Nucleic Acids Res. 1983 Dec 10;11(23):8407–8414. doi: 10.1093/nar/11.23.8407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farchaus J., Dilley R. A. Purification and partial sequence of the Mr 10,000 phosphoprotein from spinach thylakoids. Arch Biochem Biophys. 1986 Jan;244(1):94–101. doi: 10.1016/0003-9861(86)90097-4. [DOI] [PubMed] [Google Scholar]
  10. Fish L. E., Kück U., Bogorad L. Two partially homologous adjacent light-inducible maize chloroplast genes encoding polypeptides of the P700 chlorophyll a-protein complex of photosystem I. J Biol Chem. 1985 Feb 10;260(3):1413–1421. [PubMed] [Google Scholar]
  11. Fromm H., Edelman M., Koller B., Goloubinoff P., Galun E. The enigma of the gene coding for ribosomal protein S12 in the chloroplasts of Nicotiana. Nucleic Acids Res. 1986 Jan 24;14(2):883–898. doi: 10.1093/nar/14.2.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Kato A., Shimada H., Kusuda M., Sugiura M. The nucleotide sequences of two tRNAAsn genes from tobacco chloroplasts. Nucleic Acids Res. 1981 Nov 11;9(21):5601–5607. doi: 10.1093/nar/9.21.5601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kato A., Takaiwa F., Shinozaki K., Sugiura M. Location and nucleotide sequence of the genes for tobacco chloroplast tRNAArg (ACG) and tRNALeu(UAG). Curr Genet. 1985;9(5):405–409. doi: 10.1007/BF00421612. [DOI] [PubMed] [Google Scholar]
  15. Koch W., Edwards K., Kössel H. Sequencing of the 16S-23S spacer in a ribosomal RNA operon of Zea mays chloroplast DNA reveals two split tRNA genes. Cell. 1981 Jul;25(1):203–213. doi: 10.1016/0092-8674(81)90245-2. [DOI] [PubMed] [Google Scholar]
  16. Kung S. D., Lin C. M. Chloroplast promoters from higher plants. Nucleic Acids Res. 1985 Nov 11;13(21):7543–7549. doi: 10.1093/nar/13.21.7543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lerbs S., Bräutigam E., Parthier B. Polypeptides of DNA-dependent RNA polymerase of spinach chloroplasts: characterization by antibody-linked polymerase assay and determination of sites of synthesis. EMBO J. 1985 Jul;4(7):1661–1666. doi: 10.1002/j.1460-2075.1985.tb03834.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Medgyesy P., Fejes E., Maliga P. Interspecific chloroplast recombination in a Nicotiana somatic hybrid. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6960–6964. doi: 10.1073/pnas.82.20.6960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morris J., Herrmann R. G. Nucleotide sequence of the gene for the P680 chlorophyll alpha apoprotein of the photosystem II reaction center from spinach. Nucleic Acids Res. 1984 Mar 26;12(6):2837–2850. doi: 10.1093/nar/12.6.2837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ohme M., Kamogashira T., Shinozaki K., Sugiura M. Structure and cotranscription of tobacco chloroplast genes for tRNAGlu(UUC), tRNATyr(GUA) and tRNAAsp(GUC). Nucleic Acids Res. 1985 Feb 25;13(4):1045–1056. doi: 10.1093/nar/13.4.1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ohme M., Kamogashira T., Shinozoki K., Sugiura M. Locations and sequences of tobacco chloroplast genes for tRNAPro(UGG), tRNATrp, tRNAfMet and tRNAGly(GCC): the tRNAGly contains only two base-pairs in the D stem. Nucleic Acids Res. 1984 Sep 11;12(17):6741–6749. doi: 10.1093/nar/12.17.6741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohme M., Tanaka M., Chunwongse J., Shinozaki K., Sugiura M. A tobacco chloroplast DNA sequence possibly coding for a polypeptide similar to E. coli RNA polymerase beta-subunit. FEBS Lett. 1986 May 5;200(1):87–90. doi: 10.1016/0014-5793(86)80516-6. [DOI] [PubMed] [Google Scholar]
  24. Pillay D. T., Guillemaut P., Weil J. H. Nucleotide sequences of three soybean chloroplast tRNAsLeu and re-examination of bean chloroplast tRNA2Leu sequence. Nucleic Acids Res. 1984 Mar 26;12(6):2997–3001. doi: 10.1093/nar/12.6.2997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Quigley F., Weil J. H. Organization and sequence of five tRNA genes and of an unidentified reading frame in the wheat chloroplast genome: evidence for gene rearrangements during the evolution of chloroplast genomes. Curr Genet. 1985;9(6):495–503. doi: 10.1007/BF00434054. [DOI] [PubMed] [Google Scholar]
  26. Samuelsson T., Elias P., Lustig F., Axberg T., Fölsch G., Akesson B., Lagerkvist U. Aberrations of the classic codon reading scheme during protein synthesis in vitro. J Biol Chem. 1980 May 25;255(10):4583–4588. [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shinozaki K., Deno H., Kato A., Sugiura M. Overlap and cotranscription of the genes for the beta and epsilon subunits of tobacco chloroplast ATPase. Gene. 1983 Oct;24(2-3):147–155. doi: 10.1016/0378-1119(83)90074-4. [DOI] [PubMed] [Google Scholar]
  29. Shinozaki K., Deno H., Wakasugi T., Sugiura M. Tobacco chloroplast gene coding for subunit I of proton-translocating ATPase: comparison with the wheat subunit I and E. coli subunit b. Curr Genet. 1986;10(5):421–423. doi: 10.1007/BF00418416. [DOI] [PubMed] [Google Scholar]
  30. Shinozaki K., Sugiura M. Sequence of the intercistronic region between the ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit and coupling factor beta subunit gene. Nucleic Acids Res. 1982 Aug 25;10(16):4923–4934. doi: 10.1093/nar/10.16.4923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shinozaki K., Sugiura M. The nucleotide sequence of the tobacco chloroplast gene for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Gene. 1982 Nov;20(1):91–102. doi: 10.1016/0378-1119(82)90090-7. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Staden R. A computer program to search for tRNA genes. Nucleic Acids Res. 1980 Feb 25;8(4):817–825. [PMC free article] [PubMed] [Google Scholar]
  34. Stern D. B., Lonsdale D. M. Mitochondrial and chloroplast genomes of maize have a 12-kilobase DNA sequence in common. Nature. 1982 Oct 21;299(5885):698–702. doi: 10.1038/299698a0. [DOI] [PubMed] [Google Scholar]
  35. Sugita M., Shinozaki K., Sugiura M. Tobacco chloroplast tRNA(UUU) gene contains a 2.5-kilobase-pair intron: An open reading frame and a conserved boundary sequence in the intron. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3557–3561. doi: 10.1073/pnas.82.11.3557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sugita M., Sugiura M. A putative gene of tobacco chloroplast coding for ribosomal protein similar to E. coli ribosomal protein S19. Nucleic Acids Res. 1983 Mar 25;11(6):1913–1918. doi: 10.1093/nar/11.6.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Takaiwa F., Sugiura M. Nucleotide sequence of the 16S - 23S spacer region in an rRNA gene cluster from tobacco chloroplast DNA. Nucleic Acids Res. 1982 Apr 24;10(8):2665–2676. doi: 10.1093/nar/10.8.2665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takaiwa F., Sugiura M. The complete nucleotide sequence of a 23-S rRNA gene from tobacco chloroplasts. Eur J Biochem. 1982 May;124(1):13–19. doi: 10.1111/j.1432-1033.1982.tb05901.x. [DOI] [PubMed] [Google Scholar]
  39. Tohdoh N., Shinozaki K., Sugiura M. Sequence of a putative promoter region for the rRNA genes of tobacco chloroplast DNA. Nucleic Acids Res. 1981 Oct 24;9(20):5399–5406. doi: 10.1093/nar/9.20.5399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tohdoh N., Sugiura M. The complete nucleotide sequence of 16S ribosomal RNA gene from tobacco chloroplasts. Gene. 1982 Feb;17(2):213–218. doi: 10.1016/0378-1119(82)90074-9. [DOI] [PubMed] [Google Scholar]
  41. Tomioka N., Sugiura M. The complete nucleotide sequence of a 16S ribosomal RNA gene from a blue-green alga, Anacystis nidulans. Mol Gen Genet. 1983;191(1):46–50. doi: 10.1007/BF00330888. [DOI] [PubMed] [Google Scholar]
  42. Torazawa K., Hayashida N., Obokata J., Shinozaki K., Sugiura M. The 5' part of the gene for ribosomal protein S12 is located 30 kbp downstream from its 3' part in tobacco chloroplast genome. Nucleic Acids Res. 1986 Apr 11;14(7):3143–3143. doi: 10.1093/nar/14.7.3143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wilbur W. J., Lipman D. J. Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci U S A. 1983 Feb;80(3):726–730. doi: 10.1073/pnas.80.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Willey D. L., Auffret A. D., Gray J. C. Structure and topology of cytochrome f in pea chloroplast membranes. Cell. 1984 Feb;36(2):555–562. doi: 10.1016/0092-8674(84)90248-4. [DOI] [PubMed] [Google Scholar]
  45. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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