Skip to main content
Log in

Intranuclear compartmentalization of DNA-dependent RNA polymerases: association of RNA polymerase I with nucleolar organizing chromosomes

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

The compartmentalization of chromatin-bound RNA polymerases was examined with HeLa chromosomes fractionated according to their size in sucrose/glycerol gradients. There was a good correlation between the enrichment of nucleolus-organizing chromosomes, i.e., D and G group chromosomes, and the level of chromosome-associated RNA polymerase form I activity. A profile of DEAE-Sephadex chromatography of enzymes solubilized from chromosome fractions also supported the view that form I was associated with D and G group chromosomes. The amount of form I associated with unfractionated chromosomes was nearly identical to that of nuclei, indicating that this enzyme is probably quantitatively conserved even when chromatin undergoes extensive condensation. Since the chromatin-bound form I enzyme can be reactivated with heparin, it seems that this enzyme is in the initiated state, probably being bound to rDNA throughout the mitotic cycle. Thus, the absence of rRNA synthesis in mitosis is due to neither unavailability of enzyme to rDNA nor to the release of some factors necessary for transcriptional processes. Form II enzyme was associated uniformly with all chromosome fractions. Taken together, the present findings suggest that the intranuclear compartmentalization of RNA polymerases persists not only in interphase stages, but also in mitosis, during which most other nuclear proteins are released into the cytoplasm.

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

  • Bassleer R (1968) Contribution to the study of nuclear total proteins and DNA during the mitotic cycle in fibroblasts cultivated in vitro and in Ehrlich ascites cells. Histochemie 14:89–102

    Google Scholar 

  • Blatti SP, Ingles CJ, Lindell TJ, Morris PW, Weaver RF, Weinberg F, Rutter HJ (1970) Structure and regulatory properties of eukaryotic RNA polymerase. Cold Spring Harbor Quant Biol Vol 35:649–657

    Google Scholar 

  • Funaki K, Matsui S, Sasaki M (1975) Location of nucleolar organizers in animal and plant chromosomes by means of an improved N-banding technique. Chromosoma 49:357–370

    Google Scholar 

  • Gariglio P, Buss J, Green MH (1974) Sarkosyl activation of RNA polymerase activity in mitotic mouse cells. FEBS Lett 44:330–333

    Google Scholar 

  • Gilmour RS, Paul J (1969) RNA transcribed from reconstituted nucleoprotein is similar to natural RNA. J Mol Biol 40:137–139

    Google Scholar 

  • Goyanes VJ, Matsui S, Sandberg AA (1980) Chromatin organization of Chinese hamster chromosomes studied by histone-DNA crosslinking and trypsin digestion. Chromosoma 78:123–135

    Google Scholar 

  • Hannah-Alva A (1951) Localization and function of heterochromatin in Drosophila melanogaster. Adv Genet 4:87–125

    Google Scholar 

  • Heitz E (1934) Über α- and β-Heterochromatin sowie Konstanz und Bau der Chromosomeren bei Drosophila. Biol Zentralbl 54:588–609

    Google Scholar 

  • Henderson AS, Warburton D, Atwood KC (1972) Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci USA 69:3394–3398

    Google Scholar 

  • Huberman JA, Attardi G (1967) Studies of fractionated HeLa cell metaphase chromosomes. I. The chromosomal distribution of DNA complementary to 28S and 18S ribosomal RNA and to cytoplasmic messenger RNA. J Mol Biol 29:487–505

    Google Scholar 

  • Human Gene Mapping 7 (1983) In: Cytogenetics and Cell Genetics, Vol 37

  • Jaehning JA, Stewart CC, Roeder RG (1975) DNA-dependent RNA polymerase levels during the response of human peripheral lymphocytes to phytohemagglutinin. Cell 4:51–57

    Google Scholar 

  • Jamrich M, Greenleaf AL, Bautz EKF (1977a) Localization of RNA polymerase in polytene chromosomes of Drosophila melanogaster. Proc Natl Acad Sci USA 74:2079–2083

    Google Scholar 

  • Jamrich M, Greenleaf AL, Bautz FA, Bautz EKF (1977b) Functional organization of polytene chromosomes. Cold Spring Harbor Quant Biol Vol 42:389–396

    Google Scholar 

  • Johns EW, Forrester S (1969) Studies on nuclear proteins. The binding of extra acidic proteins to deoxyribonucleoprotein during the preparation of nuclear proteins. Eur J Biochem 8:547–551

    Google Scholar 

  • Johnson TC, Holland JJ (1965) Ribonucleic acid and protein synthesis in mitotic HeLa cells. J Cell Biol 27:565–574

    Google Scholar 

  • Matsui S (1974) Structural proteins associated with ribosomal cistrans in Xenopus laevis chromosomes. Exp Cell Res 88:88–94

    Google Scholar 

  • Matsui S, Sandberg AA (1983) Chromosomal mapping and ribosomal DNA (rDNA)-binding property of N-band proteins. J Cell Biol 97:137a

    Google Scholar 

  • Matsui S, Sasaki M (1973) Differential staining of nucleolus organizer in mammalian chromosomes. Nature 246:148–150

    Google Scholar 

  • Matsui S, Yoshida H, Weinfeld H, Sandberg AA (1972) Induction of prophase in interphase nuclei by fusion with metaphase cells. J Cell Biol 54:120–132

    Google Scholar 

  • Matsui S, Seon BK, Sandberg AA (1979a) Disappearance of a structural chromatin protein A24 in mitosis: Implications for molecular basis of chromatin condensation. Proc Natl Acad Sci USA 76:6386–6390

    Google Scholar 

  • Matsui S, Fuke M, Busch H (1977a) Fidelity of rDNA synthesis by nucleoli and nucleolar chromatin. Biochemistry 16:39–45

    Google Scholar 

  • Matsui S, Weinfeld H, Sandberg AA (1977b) Quantitative conservation of chromatin-bound RNA polymerase I and II in mitosis: Implications for chromosome structure. J Cell Biol 75:121a

    Google Scholar 

  • Matsui S, Weinfeld H, Sandberg AA (1978) Quantitative conservation of chromatin-bound RNA polymerases in mitosis: Compartmentalization of form I in human nucleolar organizing chromosomes. J Cell Biol 79:116a

    Google Scholar 

  • Matsui S, Weinfeld H, Sandberg AA (1979b) Quantitative conservation of chromatin-bound RNA polymerase I and II in mitosis: Implications for chromosome structure. J Cell Biol 80:451–464

    Google Scholar 

  • Morcillo G, Terre CDK, Gimenez-Martin G (1976) Nucleolar transcription during plant mitosis. In situ assay for RNA polymerase activity. Exp Cell Res 102:311–316

    Google Scholar 

  • Moore GPM, Ringertz NR (1973) Localization of DNA-dependent RNA polymerase activities in fixed human fibroblasts by autoradiography. Exp Cell Res 76:223–228

    Google Scholar 

  • Paris Conference (1972) Standardization in human cytogenetics. Birth defects Vol VIII, No 7, The National Foundation

  • Phillips DMP (1968) The binding of extra histones and protamine to deoxyribonucleoprotein. Experientia 24:668–669

    Google Scholar 

  • Prensky W, Smith HH (1964) Incorporation of 3H-arginine in chromosomes of Vicia faba. Exp Cell Res 34:525–532

    Google Scholar 

  • Prescott DM (1964) Cellular sites of RNA synthesis. In: Davidson JN, Cohn WE (eds) Progress in nucleic acids research, vol 3. Academic Press, New York and London, pp 33–57

    Google Scholar 

  • Prescott DM, Bender MA (1962) Synthesis of RNA and protein during mitosis in mammalian tissue culture cells. Exp Cell Res 26:260–268

    Google Scholar 

  • Richards GN (1974) Modifications of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA. Anal Biochem 57:369–376

    Google Scholar 

  • Roeder RG, Rutter WJ (1970) Specific nucleolar and nucleoplasmic RNA polymerases. Proc Natl Acad Sci USA 65:675–682

    Google Scholar 

  • 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

    Google Scholar 

  • Schmidt G, Thannhauser SJ (1945) A method for the determination of deoxyribonucleic acid, ribonucleic acid and phosphoproteins in animal tissues. J Biol Chem 161:83–89

    Google Scholar 

  • Tata JR, Baker B (1974) Sub-nuclear fractionation. II. Intranuclear compartmentation of transcription in vivo and in vitro. Exp Cell Res 83:125–138

    Google Scholar 

  • Wray W, Mace Jr, ML, Daskal Y (1977) Chromosome proteins and metaphase chromosome structure. In: Ts'o P (ed) The molecular biology of the mammalian genetic apparatus. Elsevier/North-Holland, pp 401–415

    Google Scholar 

  • Zylber EA, Penman S (1971a) Synthesis of 5S and 4S RNA in metaphase-arrested HeLa cells. Science 172:947–949

    Google Scholar 

  • Zylber EA, Penman S (1971b) Products of RNA polymerases in HeLa cell nuclei. Proc Natl Acad Sci USA 68:2861–2865

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Matsui, S., Sandberg, A.A. Intranuclear compartmentalization of DNA-dependent RNA polymerases: association of RNA polymerase I with nucleolar organizing chromosomes. Chromosoma 92, 1–6 (1985). https://doi.org/10.1007/BF00327238

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327238

Keywords

Navigation