Skip to main content
SpringerLink
Log in

Kinetic evidence that a discrete messenger-like RNA is formed by post-transcriptional size reduction of heterogeneous nuclear RNA

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

A discrete fraction of heterogeneous nuclear RNA, in addition to the previously described 75 S RNA, has been identified in salivary gland cells of Chironomus tentans. On the basis of its electrophoretic mobility in 1% agarose gel, it is tentatively designated as 35 S RNA. The 35 S RNA possesses distinct and characteristic labelling kinetics. After a lag of 20–45 min it emerges on the chromosomes and in the nuclear sap, but significant quantities of labelled 35 S RNA in the cytoplasm can be found only after 45–90 min of incorporation. The origin of 35 S RNA from genes manufacturing heterogeneous nuclear RNA has been established on the basis of its sensitivity against the nucleoside analogue, 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole, DRB. RNA extract derived from chromosome IV, carrying the Balbiani rings, lacks 35 S RNA. Furthermore, an essentially normal appearance of 35 S RNA was observed in glands having very small or no Balbiani rings and no detectable 75 S RNA synthesis. Both these findings are consonant with the interpretation that 35 S RNA is not generated by genes harboured in the Balbiani rings. When the continuous synthesis of chromosomal RNA is interrupted by actinomycin D, most of the 35 S RNA accumulated in the nuclear sap disappears, whereas a corresponding RNA fraction arrives in the cytoplasm. Kinetic studies as well as the chase experiment with actinomycin D suggest the existence of a kinetic relationship between nuclear sap 35 S RNA and its cytoplasmic counterpart. In addition, studies with the initiation inhibitor DRB strongly indicate that 35 S RNA derives from chromosomal RNA molecules of higher S values than 35 S, probably from RNA in the 75–100 S range, by post-transcriptional cleavages.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DRB:

5,6-dichloro-1-β-D-ribofuranosylbenzimidazole

hnRNA:

heterogeneous nuclear RNA

BR RNA:

RNA from the two major Balbiani rings, BR1 andBR2, on chromosome IV

References

  • Bachenheimer, S., Darnell, J.E.: Adenovirus-2 mRNA is transcribed as part of a high-molecular-weight precursor RNA. Proc. nat. Acad. Sci. (Wash.) 72, 4445–4449 (1975)

    Google Scholar 

  • Beermann, W.: Chromomerenkonstanz und spezifische Modifikationen der Chromosomenstruktur in der Entwicklung und Organdifferenzierung von Chironomus tentans. Chromosoma (Berl.) 5, 139–198 (1952)

    Google Scholar 

  • Beermann, W.: Ein Balbiani-Ring als Locus einer Speicheldrüsen-Mutation. Chromosoma (Berl.) 12, 1–25 (1961)

    Google Scholar 

  • Beermann, W.: Directed changes in the pattern of Balbiani ring puffing in Chironomus: Effects of sugar treatment. Chromosoma (Berl.) 41, 297–326 (1973)

    Google Scholar 

  • Bisseling, T., Berendes, H.D., Eubsen, N.H.: RNA synthesis in puff 2–48 BC after experimental induction in Drosophila hydei. Cell 8, 299–304 (1976)

    Google Scholar 

  • Daneholt, B.: Giant RNA transcript in a Balbiani ring. Nature (Lond.) 240, 229–232 (1972)

    Google Scholar 

  • Daneholt, B., Edström, J.-E., Egyházi, E., Lambert, B., Ringborg, U.: Chromosomal RNA synthesis in polytene chromosomes. Chromosoma (Berl.) 28, 399–417 (1969)

    Google Scholar 

  • Daneholt, B., Hosick, H.: Evidence for transport of 75 S RNA from a discrete chromosome region via nuclear sap to cytoplasm in Chironomus tentans. Proc. nat. Acad. Sci. (Wash.) 70, 442–446 (1973)

    Google Scholar 

  • Darnell, J.E.: The origin of mRNA and the structure of the mammalian chromosome. Harvey Lectures 69, 1–47 (1975)

    Google Scholar 

  • Davidson, E.H., Britten, R.J.: Organization, transcription and regulation in the animal genome. Quart. Rev. Biol. 48, 565–613 (1973)

    Google Scholar 

  • Doyle, D., Laufer, H.: Sources of larval salivary gland secretion in the dipteran Chironomus tentans. J. Cell Biol. 40, 61–79 (1969)

    Google Scholar 

  • Edström, J.-E.: Microextraction and microelectrophoresis for determination and analysis of nucleic acids in isolated cellular units. In: Methods in cell physiology, Vol. 1, pp. 414–447 New York: Academic Press 1964

    Google Scholar 

  • Edström, J.-E., Daneholt, B.: Sedimentation properties of the newly synthesized RNA from isolated nuclear components of Chironomus tentans salivary gland cells. J. molec. Biol. 28, 331–343 (1967)

    Google Scholar 

  • Edström, J.-E., Tanguay, R.: Cytoplasmic nucleic acid with messenger characteristics in salivary glands of Chironomus tentans. J. molec. Biol. 84, 569–583 (1974)

    Google Scholar 

  • Egyházi, E.: Actinomycin D and RNA transport. Nature (Lond.) 250, 221–223 (1974a)

    Google Scholar 

  • Egyházi, E.: A tentative initiation inhibitor of chromosomal heterogeneous RNA synthesis. J. molec. Biol. 84, 173–183 (1974b)

    Google Scholar 

  • Egyházi, E.: Processing of heterogeneous chromosomal RNA: Studies with an initiation inhibitor. Biochemistry of the Cell Nucleus, Ninth FEBS Meeting, Budapest 33, 57–62 Budapest: Akademiai Kiadó 1974

    Google Scholar 

  • Egyházi, E.: Inhibition of Balbiani ring RNA synthesis at the initiation level. Proc. nat. Acad. Sci. (Wash.) 72, 947–950 (1975)

    Google Scholar 

  • Egyházi, E.: Quantitation of turnover and export to the cytoplasm of hnRNA transcribed in the Balbiani rings. Cell 7, 507–515 (1976a)

    Google Scholar 

  • Egyházi, E.: Initiation inhibition and reinitiation of the synthesis of heterogeneous nuclear RNA in living cells. Nature (Lond.) 262, 319–321 (1976b)

    Google Scholar 

  • Egyházi, E., Daneholt, B., Edström, J.-E., Lambert, B., Ringborg, U.: Differential inhibitory effect of a substituted benzimidazole on RNA labelling in polytene chromosomes. J. Cell Biol. 47, 516–520 (1970)

    Google Scholar 

  • Firtel, R.A., Lodish, H.F.: A small nuclear precursor of messenger RNA in the cellular slime mold Dictyostelium discoideum. J. molec. Biol. 79, 295–314 (1973)

    Google Scholar 

  • Georgiev, G.P., Ryskov, A.P., Coutelle, C., Mantieva, V.L., Avakyan, E.R.: On the structure of transcriptional unit in mammalian cells. Biochim. biophys. Acta (Amst.) 259, 259–283 (1972)

    Google Scholar 

  • Greenberg, J.R., Perry, R.P.: Relative occurrence of polyadenylic acid sequences in messenger and heterogeneous nuclear RNA of L cells as determined by poly(U)-hydroxylapatit chromatography. J. molec. Biol. 72, 91–98 (1972)

    Google Scholar 

  • Grossbach, U.: Chromosomen-Aktivität und biochemische Zelldifferentierung in den Speicheldrüsen con Camptochrinomus. Chromosoma (Berl.) 28, 136–187 (1969)

    Google Scholar 

  • Grossbach, U.: Chromosome puffs and gene expression in polytene cells. Cold Spr. Harb. Symp. quant. Biol. 38, 619–627 (1973)

    Google Scholar 

  • Hollenberg, C.P.: Proportionate representation of rDNA and Balbiani ring DNA in polytene chromosomes of Chironomus tentans. Chromosoma (Berl.) 57, 185–1197 (1976)

    Google Scholar 

  • Hyman, R.W., Davidson, N.: Kinetics of the in vitro inhibition of transcription by actinomycin. J. molec. Biol. 50, 421–438 (1970)

    Google Scholar 

  • Jelinek, W., Adesnik, M., Salditt, M., Sheiness, D., Wall, R., Molloy, G., Philipson, L., Darnell, J.E.: Further evidence on the nuclear origin and transfer to the cytoplasm of polyadenylic acid sequences in mammalian cell RNA. J. molec. Biol. 75, 515–532 (1973)

    Google Scholar 

  • Lambert, B., Edström, J.-E.: Balbiani ring nucleotide sequences in cytoplasmic 75 S RNA of Chironomus tentans salivary gland cells. Mol. Biol. Reports 1, 457–464 (1974)

    Google Scholar 

  • Lee, S.Y., Mendecki, J., Brawerman, G.: A polynucleotide segment rich in polyadenylic acid in the rapidly labelled polyribosomal component of mouse sarcoma 180 ascites cells. Proc. nat. Acad. Sci. (Wash.) 68, 1331–1335 (1971)

    Google Scholar 

  • Lewin, B.: Units of transcription and translation: the relationship between heterogeneous nuclear RNA and messenger RNA. Cell 4, 11–20 (1975)

    Google Scholar 

  • Lindberg, U., Darnell, J.E.: SV40-specific RNA in the nucleus and polyribosomes of transformed cells. Proc nat. Acad. Sci. (Wash.) 65, 1089–1096 (1970)

    Google Scholar 

  • Lizardi, P.M.: The size of pulse-labeled fibroin messenger RNA. Cell 7, 239–245 (1976)

    Google Scholar 

  • Macnaughton, M., Freeman, K.B., Bishop, J.O.: A precursor to hemoglobin mRNA in nuclei of immature duck red blood cells. Cell 1, 117–125 (1974)

    Google Scholar 

  • McKnight, G.S., Schimke, R.T.: Ovalbumin messenger RNA: Evidence the initial product of transcription is the same size as polysomal ovalbumin messenger. Proc. nat. Acad. Sci. (Wash.) 71, 4327–4331 (1974)

    Google Scholar 

  • Mendecki, J., Lee, S.Y., Brawerman, G.: Characteristics of the polyadenylic acid segment associated with messenger ribonucleic acid in mouse sarcoma 180 ascites cells. Biochemistry 11, 792–798 (1972)

    Google Scholar 

  • Molloy, G.R., Jelinek, W., Salditt, M., Darnell, J.E.: Arrangement of specific oligonucleotides within poly(A) terminated HnRNA molecules. Cell 1, 43–53 (1974)

    Google Scholar 

  • Ringborg, U., Rydlander, L.: Nucleolar derived ribonucleic acid in chromosomes, nuclear sap, and cytoplasm of Chironomus tentans salivary gland cells. J. Cell Biol. 51, 355–368 (1971)

    Google Scholar 

  • Ruiz-Carrillo, A., Beato, M., Schutz, G., Feigelson, P., Allfrey, V.G.: Cell-free translation of the globin message within polydisperse high molecular weight ribonucleic acid of avian erythrocytes. Proc. nat. Acad. Sci. (Wash.) 70, 3641–3645 (1973)

    Google Scholar 

  • Rydlander, L., Edström, J.-E.: Expression of Balbiani ring genes. In: Organization and expression of the eukaryotic genome, pp. 149–156. In: Proc. of the Tenth FEBS Meeting, Paris, 1975

  • Scheer, U., Trendelenburg, M.F., Franke, W.W.: Effects of actinomycin D on the association of newly formed ribonucleo-proteins with the cistrons of ribosomal RNA in Triturus oocytes. J. Cell Biol. 65, 163–179 (1975)

    Google Scholar 

  • Scherrer, K.: Karolinska Symposia on Res. Methods in Repr. Endocrinol, 6th Symp. pp. 95–129 Geneva 1973

  • Scherrer, K., Spohr, G., Granboulan, N., Morel, G., Grosslande, J., Chezzi, C.: Nuclear and cytoplasmic messenger-like RNA and their relation to the active messenger RNA in polyribosomes of HeLa cells. Cold Spr. Harb. Symp. quant. Biol. 35, 539–554 (1970)

    Google Scholar 

  • Spohr, G., Imaizumi, T., Scherrer, K.: Synthesis and processing of nuclear precursor messenger RNA in avian erythroblasts and HeLa cells. Proc. nat. Acad. Sci. (Wash.) 71, 5009–5013 (1974)

    Google Scholar 

  • Wobus, U., Popp, S., Serfling, E., Panitz, R.: Protein synthesis in the Chironomus tentans. Molec. gen. Genet. 116, 309–321 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Histology, Karolinska Institutet, S-104 01, Stockholm 60, Sweden

    Endre Egyházi

Authors
  1. Endre Egyházi
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Egyházi, E. Kinetic evidence that a discrete messenger-like RNA is formed by post-transcriptional size reduction of heterogeneous nuclear RNA. Chromosoma 65, 137–152 (1978). https://doi.org/10.1007/BF00329466

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00329466

Keywords

Search

Navigation