Abstract
RNA polymerase I preparations purified from a rat hepatoma contained DNA topoisomerase activity. The DNA topoisomerase associated with the polymerase had an Mr of 110000, required Mg2+ but not ATP, and was recognized by anti-topoisomerase I antibodies. When added to RNA polymerase I preparations containing topoisomerase activity, anti-topoisomerase I antibodies were able to inhibit the DNA relaxing activity of the preparation as well as RNA synthesis in vitro. RNA polymerase II prepared by analogous procedures did not contain topoisomerase activity and was not recognized by the antibodies. The topoisomerase I: polymerase I complex was reversibly dissociated by column chromatography on Sephacryl S200 in the presence of 0.25 M (NH4)2SO4. Topoisomeras I was immunolocalized in the transcriptionally active ribosomal gene complex containing RNA polymerase I in situ. These data indicate that topoisomerase I and RNA polymerase I are tightly complexed both in vivo and in vitro, and suggest a role for DNA topoisomerase I in the transcription of ribosomal genes.
Similar content being viewed by others
References
Akrigg A, Cook PR (1980) DNA gyrase stimulates transcription. Nucleic Acids Res 8:845–854
Bonven BJ, Gocke E, Westergaard O (1985) A high affinity topoisomerase I binding sequence is clustered at DNAase I hypersensitive sites in Tetrahymena R-chromatin. Cell 41:541–551
Bullock P, Champoux JJ, Botchan M (1985) Association of crossover points with topoisomerase I cleavage sites: a model for nonhomologous recombination. Science 230:954–958
Champoux JJ, Dulbecco R (1972) An activity from mammalian cells that untwists superhelical DNA-A possible swivel for DNA replication. Proc Natl Acad Sci USA 69:143–146
Duceman BW, Rose KM, Jacob ST (1981) Activation of purified hepatoma RNA polymerase I by homologous protein kinase NIL J Biol Chem 256:10755–10758
Durban E, Goodenough M, Mills J, Busch H (1985) Topoisomerase I phosphorylation in vitro and in rapidly growing Novikoff hepatoma cells. EMBO J 4:2921–2926
Dynan WS, Burgess R (1981) In vitro transcription by wheat germ RNA polymerase II. J Biol Chem 256:5866–5873
Ferro AM, Olivers BM (1984) Poly(ADP-ribosylation) of DNA topoisomerase I from calf thymus. J Biol Chem 259:547–554
Fleischmann G, Pflugfelder G, Steiner EK, Javaherian K, Howard GC, Wang JC, Elgin S (1984) Drosophila DNA topoisomerase I is associated with transcriptionally active regions of the genome. Proc Natl Acad Sci USA 81:6958–6962
Garg LC, DiAngelo S, Jacob ST (1987) Role of DNA topoisomerase I in transcription of supercoiled rRNA gene. Proc Natl Acad Sci USA 84:3185–3188
Gellert M (1981) DNA Topoisomerases. Annu Rev Biochem 50:879–910
Gilmour DS, Pflugfelder G, Wang JC, Lis JT (1986) Topoisomerase I interacts with transcribed regions in Drosophila cells. Cell 44:401–407
Guldner H, Szostecki H, Vosberg H, Lakomek H, Penner E, Bautz FA (1986) Scl 70 autoantibodies from scleroderma patients recognize a 95 kDa identified as DNA topoisomerase I. Chromosoma 94:132–138
Jacob ST, Rose KM (1978) RNA polymerase and poly(A)polymerase from neoplastic tissues and cells. Methods Cancer Res 14:191–241
Jonstra-Bilen J, Ittel M, Niedergana C, Vosberg H, Mandel P (1983) DNA topoisomerase I from calf thymus is inhibited in vitro by poly(ADP-ribosylation). Eur J Biochem 136:391–396
Keller W (1975a) Characterization of purified DNA-relaxing enzyme from human tissue culture cell. Proc Natl Acad Sci USA 72:2550–2554
Keller W (1975b) Determination of the number of superhelical turns in simian virus 40 DNA by gel electrophoresis. Proc Natl Acad Sci USA 72:4876–4880
Kikuchi Y, Nash HA (1979) Nicking-closing activity associated with bacteriophage int gene product. Proc Natl Acad Sci USA 76:3760–3764
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Liu L (1984) DNA topoisomerases-Enzymes that catalyse the breaking and rejoining of DNA. CRC Crit Rev Biochem 14:1–24
Muller MT, Pfund WP, Mehta VB, Trask DR (1985) Eukaryotic type I topoisomerase is enriched in the nucleolus and catalytically active on ribosomal DNA. EMBO J 4:1237–1243
Roeder RG (1976) Eukaryotic nuclear RNA polymerase. In: Losick R, Chamberlin M (eds) RNA polymerase. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 285–389
Rose KM, Ruch PA, Morris HP, Jacob ST (1976) RNA polymerases from a rat hepatoma; partial purification and comparison of properties with corresponding liver enzymes. Biochim Biophys Acta 432:60–72
Rose KM, Bell LE, Siefkin DA, Jacob ST (1981a) A heparinsensitive nuclear protein kinase. J Biol Chem 256:7468–7477
Rose KM, Stetler DA, Jacob ST (1981b) Protein kinase activity of RNA polymerase I purified from a rat hepatoma: probable function of Mr 42000 and 24600 polypeptide. Proc Natl Acad Sci USA 78:2833–2837
Rose KM, Duceman BW, Jacob ST (1983) RNA polymerase I in hepatoma 3924A: mechanism of enhanced activity relative to liver. Adv Enzyme Regul 21:307–319
Scheer U, Rose KM (1984) Localization of RNA polymerase I in interphase cells and mitotic chromosomes by light and electron microscopic immunocytochemistry. Proc Natl Acad Sci USA 81:1431–1435
Scheer U, Hugle B, Hazan R, Rose KM (1984) Drug-induced dispersal of transcribed rRNA genes and transcriptional products: immunolocalization and silver staining of different nucleolar components in rat cells treated with 5,6-dichloro-D-ribofuranosylbenzimidazole. J Cell Biol 99:672–679
Schmitt B, Buhre Y, Vosberg H (1984) Characterization of size variants of type I DNA topoisomerase isolated from calf thymus. Eur J Biochem 144:127–134
Shastry BS (1986) 5S RNA gene specific transcription factor (TFIIIA) changes the linking number of the DNA. Biochem Biophys Res Commun 134:1086–1092
Shero JH, Bordwell B, Rothfield NF, Earnshaw WC (1986) High titers of autoantibodies to topoisomerase I (Scl-70) in sera from scleroderma patients. Science 231:737–740
Sternglanz R, DiNardo D, Voelkel KA, Nishimure Y, Hirota Y, Becherer K, Zumstein L, Wang JC (1981) Mutations of the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition. Proc Natl Acad Sci USA 78:2747–2751
Szopa J, Rose KM (1986) Cleavage of the 190-kDa subunit of DNA-dependent RNA polymerase I yields small polypeptides capable of degrading DNA. J Biol Chem 261:9022–9028
Vosberg H-P (1985) DNA topoisomerase: Enzymes that control DNA conformation. Curr Top Microbiol Immunol 114:19–102
Wang JC (1971) Interaction between DNA and an Escherichia coli protein. J Mol Biol 55:523–533
Wang JC (1985) DNA topoisomerases. Annu Rev Biochem 54:665–697
Weisbrod ST (1982) Properties of active nucleosomes as revealed by HMG 14 and 17 chromatography. Nucleic Acids Res 10:2017–2042
Zeng GC, Zannis-Hadjopoulos M, Ozer HL, Hand R (1985) Defective DNA topoisomerase I activity in a DNA ts mutant of balb/ 3T3 cells. Somatic Cell Mol Genet 11:557–569
Zhang H, Wang JC, Liu LF (1988) Involvement of DNA topoisomerase I in transcription of human ribosomal RNA genes. Proc Natl Acad Sci USA 85:1060–1064
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rose, K.M., Szopa, J., Han, FS. et al. Association of DNA topoisomerase I and RNA polymerase I: a possible role for topoisomerase I in ribosomal gene transcription. Chromosoma 96, 411–416 (1988). https://doi.org/10.1007/BF00303034
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00303034