Skip to main content
Log in

Analysis of the genes encoding the largest subunit of RNA polymerase II in Arabidopsis and soybean

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

We have cloned and sequenced the gene encoding the largest subunit of RNA polymerase II (RPB1) from Arabidopsis thaliana and partially sequenced genes from soybean (Glycine max). We have also determined the nucleotide sequence for a number of cDNA clones which encode the carboxyl terminal domains (CTDs) of RNA polymerase II from both soybean and Arabidopsis. The Arabidopsis RPB1 gene encodes a polypeptide of approximately 205 kDa, consists of 12 exons, and encompasses more than 8 kb. Predicted amino acid sequence shows eight regions of similarity with the largest subunit of other prokaryotic and eukaryotic RNA polymerases, as well as a highly conserved CTD unique to RNA polymerase II.

The CTDs in plants, like those in most other eukaryotes, consist of tandem heptapeptide repeats with the consensus amino acid sequence PTSPSYS. The portion of RPB1 which encodes the CTD in plants differs from that of RPB1 of animals and lower eukaryotes. All the plant genes examined contain 2–3 introns within the CTD encoding regions, and at least two plant genes contain an alternatively spliced intron in the 3′ untranslated region. Several clustered amino acid substitutions in the CTD are conserved in the two plant species examined, but are not found in other eukaryotes. RPB1 is encoded by a multigene family in soybean, but a single gene encodes this subunit in Arabidopsis and most other eukaryotes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. AhearnJr. JM, Bartolomei MS, West ML, Cisek LJ, Corden JL: Cloning and sequence analysis of the mouse genomic locus encoding the largest subunit of RNA polymerase II. J Biol Chem 262: 10695–10705 (1987).

    PubMed  Google Scholar 

  2. Allison LA, Moyle M, Shales M, Ingles CJ: Extensive homology among the largest subunits of eukaryotic RNA polymerases. Cell 42: 599–610 (1985).

    Article  PubMed  Google Scholar 

  3. Allison LA, Wong JK-C, Fitzpatrick VD, Moyle M, Ingles CJ: The C-terminal domain of the largest subunit of RNA polymerase II of Saccharomyces cerevisiae, Drosophila melanogaster, and mammals: a conserved structure with an essential function. Mol Cell Biol 8: 321–329 (1988).

    PubMed  Google Scholar 

  4. Allison LA, Ingles CJ: Mutations in RNA polymerase II enhance or suppress mutations in GAL4. Proc Natl Acad Sci USA 86: 2794–2798 (1989).

    PubMed  Google Scholar 

  5. Arndt KT, Styles CA, Fink GR: A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell 56: 527–537 (1989).

    Article  PubMed  Google Scholar 

  6. Benton WD, Davis RW: Screening lambda gt recombinant clones by hybridization to single plaques in situ. Science 196: 180–182 (1977).

    PubMed  Google Scholar 

  7. Bingham ET, Cutter GL, Beversdorf WD: Creating genetic variability: tissue culture and chromosome manipulation. In: Hill LD (ed.) World Soybean Research. Interstate Printers and Publishers, Inc., Danville IL (1976).

    Google Scholar 

  8. Bartolomei MS, Handen NF, Cullen CR, Corden JL: Genetic analysis of the repetitive carboxyl-terminal domain of the largest subunit of mouse RNA polymerase II. Mol Cell Biol 8: 330–339 (1988).

    PubMed  Google Scholar 

  9. Bernstein P, Pletz SW, Ross J: The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro. Mol Cell Biol 9: 659–670 (1989).

    PubMed  Google Scholar 

  10. Biggs J, Searles LL, Greenleaf AL: Structure of the eukaryotic transcription apparatus: features of the gene for the largest subunit of Drosophila RNA polymerase II. Cell 42: 611–621 (1985).

    Article  PubMed  Google Scholar 

  11. Bird DM, Riddle DR: Molecular cloning and sequence of ama-1, the gene encoding the largest subunit of Caenorhabditis elegans RNA polymerase II. Mol Cell Biol 9: 4119–4130 (1989).

    PubMed  Google Scholar 

  12. Brandl CJ, Struhl K: Yeast GCN4 transcriptional activator protein interacts with RNA polymerase II in vitro. Proc Natl Acad Sci USA 86: 2652–2656 (1989).

    PubMed  Google Scholar 

  13. Breitbart RD, Andreadis A, Nadal-Ginard B: Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Ann Rev Biochem 56: 467–495 (1987).

    Article  PubMed  Google Scholar 

  14. Brown J: A catalogue of splice junction and putative branch point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).

    PubMed  Google Scholar 

  15. Broyles SS, Moss R: Homology between RNA polymerases of poxviruses, prokaryotes, and eukaryotes: Nucleotide sequence and transcriptional analysis of vaccinia virus genes encoding the 147-kDa and 22-kDa subunits. Proc Natl Acad Sci USA 83: 3141–3145 (1986).

    PubMed  Google Scholar 

  16. Cadena DL, Dahmus ME: Messenger RNA synthesis in mammalian cells is catalyzed by the phosphorylated form of RNA polymerase II. J Biol Chem 262: 12468–12474 (1987).

    PubMed  Google Scholar 

  17. Cisek LJ, Corden JL: Phosphorylation of RNA polymerase by the murine homologue of the cell-cycle control protein cdc2. Nature 339: 679–684 (1989).

    Article  PubMed  Google Scholar 

  18. Clark J, Clissold PM, Shawi RA, Beattie P, Bishop J: Structure of mouse major urinary protein genes: different splicing configurations in the 3′-non-coding region. EMBO J 3: 1045–1052 (1984).

    PubMed  Google Scholar 

  19. Corden JL, Cadena DL, AhearnJr JM, Dahmus ME: A unique structure at the carboxyl terminus of the largest subunit of eukaryotic RNA polymerase II. Proc Natl Acad Sci USA 82: 7934–7938 (1985).

    PubMed  Google Scholar 

  20. Cornelissen AWCA, Evers R, Kock J: Structure and sequence of genes encoding subunits of eukaryotic RNA polymerases. In: Maclean M (ed.) Oxford Surveys on Eukaryotic Genes, vol. 5, pp. 91–131. Oxford University Press, New York (1988).

    Google Scholar 

  21. Dahmus ME, Kedinger C: Transcription of Adenovirus-2 major late promoter inhibited by monoclonal antibody directed against RNA polymerases IIO and IIA. J Biol Chem 258: 2303–2307 (1983).

    PubMed  Google Scholar 

  22. Dale RMK, McClure BA, Houchins JP: A rapid singlestranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18s rDNA. Plasmid 13: 31–40 (1985).

    PubMed  Google Scholar 

  23. Dellaporta SL, Woods J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1(4): 19–21 (1983).

    Google Scholar 

  24. Evers R, Hammer A, Kock J, Jess W, Borst P, Memet S, Cornelissen AWCA: Trypanosoma brucei contains two RNA polymerase II largest subunit genes with an altered C-terminal domain. Cell 56: 585–597 (1989).

    Article  PubMed  Google Scholar 

  25. Evers R, Hammer A, Cornelissen AWCA: Unusual C-terminal domain of the largest subunit of RNA polymerase II of Crithidia fasciculata. Nucl Acids Res 17: 3403–3413 (1989).

    PubMed  Google Scholar 

  26. Frischauf A, Lehrach H, Poustka A, Murray N: Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170: 827–842 (1983).

    PubMed  Google Scholar 

  27. Guilfoyle TJ: Auxin-induced DNA dependent RNA polymerase activities in mature soybean hypocotyl. Biochemistry 19: 6112–6118 (1980).

    PubMed  Google Scholar 

  28. Guilfoyle TJ: DNA-dependent RNA polymerases of plants and lower eukaryotes. In: Jacobs ST (ed.) Enzymes of Nucleic Acid Synthesis and Modification, vol. II, pp. 1–42. CRC Press, Boca Raton, FL (1983).

    Google Scholar 

  29. Guilfoyle TJ: A protein kinase from wheat germ that phosphorylates the largest subunit of RNA polymerase II. Plant Cell 1: 827–836 (1989).

    Article  PubMed  Google Scholar 

  30. Hagen G, Kleinschmidt A, Guilfoyle TJ: Auxinregulated gene expression in intact soybean hypocotyl and excised hypocotyl sections. Planta 162: 147–153 (1984).

    Google Scholar 

  31. Hagen G, Guilfoyle TH: Rapid induction of selective transcription by auxins. Mol Cell Biol 5: 1197–1203 (1985).

    PubMed  Google Scholar 

  32. Hu N, Messing J: The making of strand-specific M13 probes. Gene 17: 271–277 (1982).

    Article  PubMed  Google Scholar 

  33. Huynh TV, Young RA, Davis RW: Constructing and screening cDNA libraries in lambda gt10. In: Glover DM (ed.) DNA Cloning Techniques: A Practical Approach, vol. 1, pp. 49–78. IRL Press, London (1985).

    Google Scholar 

  34. Ichiyoshi Y, Endo H, Yamamoto M: Length polymorphism in the 3′ noncoding region rat hepatic α mRNAs. Biochim Biophys Acta 910: 43–51 (1987).

    PubMed  Google Scholar 

  35. Ingelbrecht ILW, Herman LMF, Dekeyser RA, Van Montagu MC, Depicker AG: Different 3′ end regions strongly influence the level of gene expression in plant cells. Plant Cell 1: 671–680 (1989).

    Article  PubMed  Google Scholar 

  36. Johnson DA, Gautsch JW, Sportsman JR, Elder JH: Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech 1: 3–8 (1984).

    Article  Google Scholar 

  37. Jokerst RS, Weeks JR, Zehring WA, Greenleaf AL: Analysis of the gene encoding the largest subunit of RNA polymerase II in Drosophila. Mol Gen Genet 215: 266–275 (1989).

    PubMed  Google Scholar 

  38. Joshi CP: An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl Acids Res 15: 6643–6653 (1987).

    PubMed  Google Scholar 

  39. Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl Acids Res 15: 9627–9640 (1987).

    PubMed  Google Scholar 

  40. Kim W-Y, Dahmus ME: Immunological analysis of mammalian RNA polymerase II subspecies: stability and relative in vivo concentration. J Biol Chem 261: 14219–14225 (1986).

    PubMed  Google Scholar 

  41. Kim W-Y, Dahmus ME: The major late promoter of adenovirus-2 is accurately transcribed by RNA polymerases IIO, IIA, and IIB. J Biol Chem 264: 3169–3176 (1989).

    PubMed  Google Scholar 

  42. Kislev N, Rubinstein I: Utility of ethidium bromide in extraction from whole plants of high molecular weight maize DNA. Plant Phys 66: 1140–1143 (1980).

    Google Scholar 

  43. Lee JM, Greenleaf AL: A protein kinase that phophorylates the C-terminal repeat domain of the largest subunit of RNA polymerase II. Proc Natl Acad Sci USA 86: 3624–3628 (1989).

    PubMed  Google Scholar 

  44. Loenen WAM, Blattner RR: Lambda Charon vectors (Ch32, 33, 34, and 35) adapted for DNA cloning in recombination deficient hosts. Gene 26: 171–179 (1983).

    Article  PubMed  Google Scholar 

  45. McMaster GK, Carmichael GG: Analysis of single and double-stranded nucleic acid on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74: 4835–4838 (1977).

    PubMed  Google Scholar 

  46. Maniatis T, Fritsch ER, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  47. Meyerowitz E. Arabidopsis thaliana. Ann Rev Genet 21: 93–111 (1987).

    Article  PubMed  Google Scholar 

  48. Nonet M, Sweetser D, Young RA: Eucaryotic RNA polymerase conditional mutant that rapidly ceases mRNA synthesis. Mol Cell Biol 7: 1602–1611 (1987).

    PubMed  Google Scholar 

  49. Nonet M, Sweetser D, Young RA: Functional redundancy and structural polymorphism in the large subunit of RNA polymerase II. Cell 50: 909–915 (1987).

    Article  PubMed  Google Scholar 

  50. Olszewski N, Martin F, Ausubel F: Specialized binary vector for plant transformation: expression of Arabidopsis thaliana AHAS gene in Nicotiana tabacum. Nucl Acids Res 16: 10765–10782 (1988).

    PubMed  Google Scholar 

  51. Ovchinnikov YA, Monastyrskaya GS, Gubanov VV, Guryev SO, Salomatina IS, Shuvaeva TM, Lipkin VM, Sverdlov ED: The primary structure of E. coli RNA polymerase. Nucleotide sequence of the rpoC gene and amino acid sequence of the β′-subunit. Nucl Acids Res 10: 4035–4044 (1982).

    PubMed  Google Scholar 

  52. Padgett RA, Grabowski PJ, Konarska MM, Seiler S, Sharp PA: Splicing of messenger RNA precursors. Ann Rev Biochem 55: 1119–1150 (1986).

    Article  PubMed  Google Scholar 

  53. Price DH, Sluder AE, Greenleaf AL: Dynamic interaction between a Drosophila transcription factor and RNA polymerase II. Mol Cell Biol 9: 1465–1475 (1989).

    PubMed  Google Scholar 

  54. Reed KC, Mann DA: Rapid transfer of DNA from agarose gels to nylon membrane. Nucl Acids Res 13: 7207–7221 (1985).

    PubMed  Google Scholar 

  55. Sanger R, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    PubMed  Google Scholar 

  56. Searles LL, Jokerst RS, Bingham PM, Voelker RA, Greenleaf AL: Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P-element transposon tagging. Cell 31: 585–592 (1982).

    Article  PubMed  Google Scholar 

  57. Smith JL, Levin JR, Ingles CJ, Agabian N: In trypanosomes the homolog of the largest subunit of RNA polymerase II is encoded by two genes and has a highly unusual C-terminal domain structure. Cell 56: 815–827 (1989).

    Article  PubMed  Google Scholar 

  58. Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).

    PubMed  Google Scholar 

  59. Thompson NE, Steinberg TH, Aronson DB, Burgess RR: Inhibition of in vivo and in vitro transcription by monoclonal antibodies prepared against wheat germ RNA polymerase II that react with the heptapeptide repeat of eukaryotic RNA polymerase II. J Biol Chem 264: 11511–11520 (1989).

    PubMed  Google Scholar 

  60. Wereneke JM, Ogren WL: Alternative mRNA splicing generates the two ribulosebisphosphate carboxylase/oxygenase activase polypeptides in spinach and Arabidopsis. Plant Cell 1: 815–825 (1989).

    Article  PubMed  Google Scholar 

  61. Young R, Davis R: Yeast RNA polymerase II genes: isolation with antibody probes. Science 222: 778–782 (1983).

    PubMed  Google Scholar 

  62. Zehring WA, Lee JM, Weeks JR, Jokerst RS, Greenleaf AL: The C-terminal repeat domain of RNA polymerase II largest subunit is essential in vivo but is not required for accurate transcription initiation in vitro. Proc Natl Acad Sci USA 85: 3698–3702 (1988).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dietrich, M.A., Prenger, J.P. & Guilfoyle, T.J. Analysis of the genes encoding the largest subunit of RNA polymerase II in Arabidopsis and soybean. Plant Mol Biol 15, 207–223 (1990). https://doi.org/10.1007/BF00036908

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00036908

Key words

Navigation