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.
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
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
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
Gariglio P, Buss J, Green MH (1974) Sarkosyl activation of RNA polymerase activity in mitotic mouse cells. FEBS Lett 44:330–333
Gilmour RS, Paul J (1969) RNA transcribed from reconstituted nucleoprotein is similar to natural RNA. J Mol Biol 40:137–139
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
Hannah-Alva A (1951) Localization and function of heterochromatin in Drosophila melanogaster. Adv Genet 4:87–125
Heitz E (1934) Über α- and β-Heterochromatin sowie Konstanz und Bau der Chromosomeren bei Drosophila. Biol Zentralbl 54:588–609
Henderson AS, Warburton D, Atwood KC (1972) Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci USA 69:3394–3398
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
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
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
Jamrich M, Greenleaf AL, Bautz FA, Bautz EKF (1977b) Functional organization of polytene chromosomes. Cold Spring Harbor Quant Biol Vol 42:389–396
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
Johnson TC, Holland JJ (1965) Ribonucleic acid and protein synthesis in mitotic HeLa cells. J Cell Biol 27:565–574
Matsui S (1974) Structural proteins associated with ribosomal cistrans in Xenopus laevis chromosomes. Exp Cell Res 88:88–94
Matsui S, Sandberg AA (1983) Chromosomal mapping and ribosomal DNA (rDNA)-binding property of N-band proteins. J Cell Biol 97:137a
Matsui S, Sasaki M (1973) Differential staining of nucleolus organizer in mammalian chromosomes. Nature 246:148–150
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
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
Matsui S, Fuke M, Busch H (1977a) Fidelity of rDNA synthesis by nucleoli and nucleolar chromatin. Biochemistry 16:39–45
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
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
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
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
Moore GPM, Ringertz NR (1973) Localization of DNA-dependent RNA polymerase activities in fixed human fibroblasts by autoradiography. Exp Cell Res 76:223–228
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
Prensky W, Smith HH (1964) Incorporation of 3H-arginine in chromosomes of Vicia faba. Exp Cell Res 34:525–532
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
Prescott DM, Bender MA (1962) Synthesis of RNA and protein during mitosis in mammalian tissue culture cells. Exp Cell Res 26:260–268
Richards GN (1974) Modifications of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA. Anal Biochem 57:369–376
Roeder RG, Rutter WJ (1970) Specific nucleolar and nucleoplasmic RNA polymerases. Proc Natl Acad Sci USA 65:675–682
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
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
Tata JR, Baker B (1974) Sub-nuclear fractionation. II. Intranuclear compartmentation of transcription in vivo and in vitro. Exp Cell Res 83:125–138
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
Zylber EA, Penman S (1971a) Synthesis of 5S and 4S RNA in metaphase-arrested HeLa cells. Science 172:947–949
Zylber EA, Penman S (1971b) Products of RNA polymerases in HeLa cell nuclei. Proc Natl Acad Sci USA 68:2861–2865
Author information
Authors and Affiliations
Rights 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
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00327238