Skip to main content
Log in

Primary structure and functional aspects of the gene coding for the second-largest subunit of RNA polymerase III of Drosophila

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

We have cloned and sequenced the gene coding for the second-largest subunit of RNA polymerase III of Drosophila melanogaster (DmRP135). The gene, interrupted by two introns of 62 and 59 bp, respectively, codes for an mRNA of 3.6 kb. As for other housekeeping genes transcription initiates at several sites (between positions −98 and −76) none of which is preceded by a clear TATA sequence. The deduced polypeptide consists of 1129 amino acids with an aggregate molecular weight of 128 kDa. The protein sequence features the same regions of similarity as observed for the corresponding subunits of RNA polymerase II of Drosophila and yeast and the Escherichia coli β subunit. As in the second-largest subunit of RNA polymerase II there is a zinc-binding motif which is absent in the β subunit of E. coli. Antibodies directed against a fusion protein expressing 164 amino acids of the (DmRP135) polypeptide cross-react with the second-largest subunit of RNA polymerase III of yeast and generate a distinct banding pattern on Drosophila polytene chromosomes distinguishable from that obtained with anti-RNA polymerase II antibodies.

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

  • Bark C, Weller P, Zabielki J, Janson L, Petterson U (1987) A distant enhancer element is required for pol III transcription of a U6 RNA gene. Nature 328:356–359

    Google Scholar 

  • Bentley D, Groudine M (1988) Sequence requirements for premature termination of transcription in the human c-myc gene. Cell 53:245–256

    Google Scholar 

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

    Google Scholar 

  • Berg JM (1986) Potential metal-binding domains in nucleic acid binding proteins. Science 232:485–487

    Google Scholar 

  • Berghöfer B, Kröckel L, Körtner C, Truss M, Schallenberg J, Klein A (1988) Relatedness of archaebacterial RNA polymerase core subunits to their eubacterial and eukaryotic equivalents. Nucleic Acids Res 16:8113–8128

    Google Scholar 

  • Biggin MD, Gibson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965

    Google Scholar 

  • Birnstiel ML, Busslinger M, Strub K (1985) Transcription termination and 3′processing: the end is in site! Cell 41:349–359

    Google Scholar 

  • Carbon P, Murgo S, Ebel J-P, Krol A, Tebb G, Mattaj I (1987) A common octamer motif binding protein is involved in the transcription of U6 snRNA by RNA polymerase III and U2 snRNA polymerase II. Cell 51:71–79

    Google Scholar 

  • Chung J, Sussman DJ, Zeller R, Leder P (1987) The c-myc gene encodes superimposed RNA polymerase II and III promoters. Cell 51:1001–1008

    Google Scholar 

  • Cornelissen AWCA, Evers R, Köck J (1988) Structure and sequence of genes encoding subunits of eukaryotic RNA polymerases. In: Oxford Surveys on Eukaryotic Genes, vol 5, p 91

    Google Scholar 

  • Dahlberg JE, Lund E (1987) The genes and transcription of the major small nuclear RNAs. In: Birnstiel M (ed) Structure and functions of small nuclear ribonucleoprotein particles. Springer, Berlin Heidelberg New York p 38

    Google Scholar 

  • Falkenburg D, Dworniczak B, Faust DM, Bautz EKE (1987) RNA polymerase II of Drosophila — relation of its 140000 Mr subunit of the β subunit of E. coli RNA polymerase. J Mol Biol 195:929–937

    Google Scholar 

  • Faust DM, Renkawitz-Pohl R, Falkenburg D, Gasch A, Bialojan S, Young RA, Bautz EKF (1986) Cloning and indentification of the gene coding for the 140-kd subunit of Drosophila RNA polymerase II. EMBO J 5:741–746

    Google Scholar 

  • Folk WR (1988) Changing direction in pol III transcription. Genes Dev 2:373–375

    Google Scholar 

  • Garcia AD, O'Connell AM, Sharp SJ (1987) Formation of an active transcription complex in the Drosophila melanogaster 5S RNA gene is dependent on an upstream region. Mol Cell Biol 7:2046–2051

    Google Scholar 

  • Grachev MA, Lukhtanov EA, Mustaev AA, Richter VA, Rabinov IV, Skoblov YS, Abdukayumov MN (1987a) Localization of lysine residues nearby the site of initiating substrate binding of E. coli RNA polymerase. Bioorg Khim 13:552–555

    Google Scholar 

  • Grachev MA, Kolocheva TI, Lukhtanov EA, Mustaev AA (1987b) Studies on the functional topography of Escherichia coli RNA polymerase. Highly selective affinity labelling by analogues of initiating substrates. Eur J Biochem 163:113–121

    Google Scholar 

  • Gundelfinger ED (1983) Interaction of nucleic acids with DNA-dependent RNA polymerases of Drosophila. FEBS Lett 157:133–138

    Google Scholar 

  • Jamrich M, Greenleaf AL, Bautz EKE (1977) Functional organization of polytene chromosomes. Proc Natl Acad Sci USA 74:2079–2083

    Google Scholar 

  • Krol A, Carbon P, Ebel J-P, Appel B (1987) Xenopus tropicalis U6 snRNA genes transcribed by pol III contain the upstream promoter elements used by pol II dependent U snRNA genes. Nucleic Acids Res 15:2463–2478

    Google Scholar 

  • Kunkel RG, Maser RL, Calvet JP, Pederson T (1986) U6 snRNA is transcribed by RNA polymerase III. Proc Natl Acad Sci USA 83:8575–8579

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Leffers H, Gropp F, Lottspeich F, Zillig W, Garrett RA (1989) Sequence, organisation, transcription and evolution of RNA polymerase subunit genes from the archaebacterial extreme halophiles Halobacterium halobium and Halococcus morrhnae. J Mol Biol 206:1–17

    Google Scholar 

  • Lemeur M, Glanville N, Mandel JL, Gerlinger P, Palmito R, Chambon P (1981) The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation. Cell 23:561–571

    Google Scholar 

  • Lisitsyn NA, Monastyrskaya GS, Sverdlov ED (1988) Genes coding for RNA polymerase β subunits in bacteria. Eur J Biochem 177:363–369

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook S (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Mattaj IW, Dathan NA, Parry HD, Carbon P, Krol A (1988) Changing the RNA polymerase specificity of U snRNA gene promoters. Cell 55:435–442

    Google Scholar 

  • Melton DW, McEwan C, McKie AB, Reid AM (1986) Expression of the mouse HPRT gene: deletional analysis of the promoter region of an X-chromosome linked housekeeping gene. Cell 44:319–328

    Google Scholar 

  • Mémet SJ, Gouy M, Marck C, Sentenac A, Buhler J-M (1988) RPA190, the gene coding for the largest subunit of yeast RNA polymerase A. J Biol Chem 263:2830–2839

    Google Scholar 

  • Mount SM (1982) A catalogue of splice junction sequences. Nucleic Acids Res 10:5495–5513

    Google Scholar 

  • Ohme M, Tanaka M, Chunwongse J, Shinzaki K, Sugiura M (1986) A tobacco chloroplast DNA sequence possibly coding for a polypeptide similar to E. coli RNA polymerase β subunit. FEBS Lett 200:87–89

    Google Scholar 

  • Patel DD, Pickup DJ (1989) The second-largest subunit of poxvirus RNA polymerase is similar to the corresponding subunits of prokaryotic and eukaryotic RNA polymerase. J Virol 63:1076–1086

    Google Scholar 

  • Rappaport J, Cho K, Saltzman A, Prenger J, Golomb M, Weinman R (1988) Transcription elongation factor SII interacts with a domain of the large subunit of human RNA pol II. Mol Cell Biol 8:3136–3142

    Google Scholar 

  • Reddy R, Henning D, Das G, Harles M, Wright D (1987) The capped U6 snRNA is transcribed by RNA polymerase III. J Biol Chem 262:75–81

    Google Scholar 

  • Riva M, Schäffner AR, Sentenac A, Hartmann GR, Mustaev AA, Zaychikov EF, Grachev MA (1987) Active site labeling of the RNA polymerase A, B, and C from yeast. J Biol Chem 262:14377–14380

    Google Scholar 

  • Rüther U, Müller-Hill B (1983) Easy identification of cDNA clones. EMBO J 2:1791–1794

    Google Scholar 

  • Sajjadi FG, Miller RC Jr, Spiegelman GB (1987) Identification of sequences in the 5′ flanking region of Drosophila melanogaster tRNA4 Val gene modulate its transcription in vitro. Mol Gen Genet 206:276–284

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sentenac A (1985) Eukaryotic RNA polymerases. CRC Crit Rev Biochem 18:31–90

    Google Scholar 

  • Sollner-Webb B (1988) Surprises in polymerase III transcription. Cell 52:153–154

    Google Scholar 

  • Sweetser B, Nonet M, Young RA (1987) Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. Proc Natl Acad Sci USA 84:1192–1196

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1981) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets, procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Williams IF, Mason PJ (1985) Hybridisation in the analysis of RNA. In: Harres BD, Heggins SJ (eds) Nucleic acid hybridisation. IRL Press, Oxford, UK pp 139–159

    Google Scholar 

  • Young RA, Davis RW (1983) Yeast RNA polymerase II genes: Isolation with antibody probes. Science 222:778–782

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J.A. Campos-Ortega

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kontermann, R., Sitzler, S., Seifarth, W. et al. Primary structure and functional aspects of the gene coding for the second-largest subunit of RNA polymerase III of Drosophila . Mol Gen Genet 219, 373–380 (1989). https://doi.org/10.1007/BF00259609

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00259609

Key words

Navigation