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PMID: 7536746 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Transcription-dependent redistribution of the large subunit of RNA polymerase II to discrete nuclear domains.

The Journal of cell biology ·Vol. 129 ·No. 2 ·1995-04-00 ·Pages 287-98

Bregman DB, Du L, van der Zee S, Warren SL

Abstract

A subpopulation of the largest subunit of RNA polymerase II (Pol II LS) is located in 20-50 discrete subnuclear domains that are closely linked to speckle domains, which store splicing proteins. The speckle-associated fraction of Pol II LS is hyperphosphorylated on the COOH-terminal domain (CTD), and it is highly resistant to extraction by detergents. A diffuse nucleoplasmic fraction of Pol II LS is relatively hypophosphorylated on the CTD, and it is easily extracted by detergents. In transcriptionally active nuclei, speckle bound hyperphosphorylated Pol II LS molecules are distributed in irregularly shaped speckle domains, which appear to be interconnected via a reticular network. When transcription is inhibited, hyperphosphorylated Pol II LS and splicing protein SC35 accumulate in speckle domains, which are transformed into enlarged, dot-like structures lacking interconnections. When cells are released from transcriptional inhibition, Pol IIO and SC35 redistribute back to the interconnected speckle pattern of transcriptionally active cells. The redistribution of Pol II and SC35 is synchronous, reversible, and temperature dependent. It is concluded that: (a) hyperphosphorylation of Pol II LS's CTD is a better indicator of its tight association to discrete subnuclear domains than its transcriptional activity; (b) during states of transcriptional inhibition, hyperphosphorylated Pol II LS can be stored in enlarged speckle domains, which under the light microscope appear to coincide with the storage sites for splicing proteins; and (c) Pol II and splicing proteins redistribute simultaneously according to the overall transcriptional activity of the nucleus.

MeSH Terms
Amanitins/pharmacology Amino Acid Sequence Animals Antibodies, Monoclonal Cell Line Cell Nucleus/enzymology,metabolism Dichlororibofuranosylbenzimidazole/pharmacology Epitopes/analysis Liver/enzymology Molecular Sequence Data Nuclear Proteins/metabolism Phosphorylation RNA Polymerase II/analysis,antagonists & inhibitors,immunology,isolation & purification,metabolism Rats Ribonucleoproteins Spliceosomes/metabolism Temperature Transcription, Genetic/drug effects
Chemicals
Amanitins Antibodies, Monoclonal Epitopes Nuclear Proteins Ribonucleoproteins Dichlororibofuranosylbenzimidazole RNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bregman D B
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Du L
van der Zee S
Warren S L
References (67)
67 references, click to expand
  1. The nonphosphorylated form of RNA polymerase II preferentially associates with the preinitiation complex.
    Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10004-8 PMID: 1946417
  2. Discrete nuclear domains of poly(A) RNA and their relationship to the functional organization of the nucleus.
    J Cell Biol. 1991 Dec;115(5):1191-202 PMID: 1720123
  3. Visualization of RNA transcription and processing.
    Semin Cell Biol. 1991 Apr;2(2):131-40 PMID: 1720682
  4. Macromolecular domains within the cell nucleus.
    Annu Rev Cell Biol. 1993;9:265-315 PMID: 8280462
  5. Splicing of Balbiani ring 1 gene pre-mRNA occurs simultaneously with transcription.
    Cell. 1994 Jan 14;76(1):183-92 PMID: 8287477
  6. RNA polymerase II is aberrantly phosphorylated and localized to viral replication compartments following herpes simplex virus infection.
    J Virol. 1994 Feb;68(2):988-1001 PMID: 8289400
  7. Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors.
    J Cell Biol. 1994 Feb;124(3):249-60 PMID: 8294510
  8. Phosphorylation of the RNA polymerase II largest subunit during heat shock and inhibition of transcription in HeLa cells.
    J Cell Physiol. 1994 Mar;158(3):417-26 PMID: 8126066
  9. Cytostellin distributes to nuclear regions enriched with splicing factors.
    J Cell Sci. 1994 Mar;107 ( Pt 3):387-96 PMID: 8006060
  10. Phosphorylation of RNA polymerase II C-terminal domain and transcriptional elongation.
    Nature. 1994 Jul 7;370(6484):75-7 PMID: 8015613
  11. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  12. Specific inhibition of nuclear RNA polymerase II by alpha-amanitin.
    Science. 1970 Oct 23;170(3956):447-9 PMID: 4918258
  13. Inhibition of RNA synthesis by actinomycin D: characteristic dose-response of different RNA species.
    J Cell Physiol. 1970 Oct;76(2):127-39 PMID: 5500970
  14. Localisation of rapidly and slowly labelled nuclear RNA as visualized by high resolution autoradiography.
    Exp Cell Res. 1971 Jul;67(1):129-41 PMID: 4105558
  15. Localization and characterization of newly synthesized nuclear RNA in isolate rat hepatocytes.
    Exp Cell Res. 1976 Apr;99(1):155-64 PMID: 944140
  16. Action of dichlorobenzimidazole riboside on RNA synthesis in L-929 and HeLa cells.
    J Cell Biol. 1976 May;69(2):229-40 PMID: 1262389
  17. The inhibition by DRB (5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole) of hnRNA and mRNA production in HeLa cells.
    Cell. 1976 Nov;9(3):473-80 PMID: 1086720
  18. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  19. The ultrastructural visualization of nucleolar and extranucleolar RNA synthesis and distribution.
    Int Rev Cytol. 1980;65:255-99 PMID: 6156137
  20. Monoclonal antibody directed against RNA polymerase II of Drosophila melanogaster.
    Mol Gen Genet. 1980;180(1):193-9 PMID: 6777631
  21. Phosphorylation of eukaryotic DNA-dependent RNA polymerase. Identification of calf thymus RNA polymerase subunits phosphorylated by two purified protein kinases, correlation with in vivo sites of phosphorylation in HeLa cell RNA polymerase II.
    J Biol Chem. 1981 Apr 10;256(7):3332-9 PMID: 6937464
  22. Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease.
    Proc Natl Acad Sci U S A. 1981 May;78(5):2737-41 PMID: 6789322
  23. Immunoelectron microscopic localization of snRNPs.
    Biol Cell. 1983;49(1):1-10 PMID: 6230127
  24. Distribution of the 70K U1 RNA-associated protein during interphase and mitosis. Correlation with other U RNP particles and proteins of the nuclear matrix.
    J Cell Sci. 1986 Dec;86:173-90 PMID: 2958489
  25. Mapping of B cell epitopes on small nuclear ribonucleoproteins that react with human autoantibodies as well as with experimentally-induced mouse monoclonal antibodies.
    J Immunol. 1989 Oct 15;143(8):2560-6 PMID: 2477448
  26. The transition of RNA polymerase II from initiation to elongation is associated with phosphorylation of the carboxyl-terminal domain of subunit IIa.
    J Biol Chem. 1989 Nov 25;264(33):19621-9 PMID: 2584185
  27. Higher order nuclear organization: three-dimensional distribution of small nuclear ribonucleoprotein particles.
    Proc Natl Acad Sci U S A. 1990 Jan;87(1):147-51 PMID: 2136950
  28. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus.
    Nature. 1990 Feb 1;343(6257):437-41 PMID: 2137203
  29. RNA polymerase B (II) and general transcription factors.
    Annu Rev Biochem. 1990;59:711-54 PMID: 2197989
  30. Phosphorylation of RNA polymerase IIA occurs subsequent to interaction with the promoter and before the initiation of transcription.
    J Biol Chem. 1990 Aug 5;265(22):13165-73 PMID: 2376591
  31. U1 and U2 small nuclear RNAs are present in nuclear speckles.
    Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):305-8 PMID: 1530887
  32. A normally masked nuclear matrix antigen that appears at mitosis on cytoskeleton filaments adjoining chromosomes, centrioles, and midbodies.
    J Cell Biol. 1992 Feb;116(4):977-87 PMID: 1734026
  33. Transcription-dependent colocalization of the U1, U2, U4/U6, and U5 snRNPs in coiled bodies.
    J Cell Biol. 1992 Apr;117(1):1-14 PMID: 1532583
  34. Preferential distribution of active RNA polymerase II molecules in the nuclear periphery.
    Gene Expr. 1991 Apr;1(1):61-70 PMID: 1668144
  35. Cytostellin: a novel, highly conserved protein that undergoes continuous redistribution during the cell cycle.
    J Cell Sci. 1992 Oct;103 ( Pt 2):381-8 PMID: 1478941
  36. Initiation of transcription by RNA polymerase II: a multi-step process.
    Prog Nucleic Acid Res Mol Biol. 1993;44:67-108 PMID: 8434126
  37. Higher level organization of individual gene transcription and RNA splicing.
    Science. 1993 Feb 26;259(5099):1326-30 PMID: 8446901
  38. A three-dimensional view of precursor messenger RNA metabolism within the mammalian nucleus.
    Science. 1993 Feb 26;259(5099):1330-5 PMID: 8446902
  39. Visualization of focal sites of transcription within human nuclei.
    EMBO J. 1993 Mar;12(3):1059-65 PMID: 8458323
  40. Association of individual hnRNP proteins and snRNPs with nascent transcripts.
    J Cell Biol. 1993 Apr;121(2):219-28 PMID: 8468343
  41. Positive patches and negative noodles: linking RNA processing to transcription?
    Trends Biochem Sci. 1993 Apr;18(4):117-9 PMID: 8493720
  42. Phosphorylation of C-terminal domain of RNA polymerase II is not required in basal transcription.
    Nature. 1993 May 27;363(6427):371-4 PMID: 8497323
  43. Ultrastructural localization of transcription sites and of RNA distribution during the cell cycle of synchronized CHO cells.
    Exp Cell Res. 1978 May;113(2):327-37 PMID: 95675
  44. Splice site selection, rate of splicing, and alternative splicing on nascent transcripts.
    Genes Dev. 1988 Jun;2(6):754-65 PMID: 3138163
  45. 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits transcription elongation by RNA polymerase II in vitro.
    J Biol Chem. 1989 Feb 5;264(4):2250-7 PMID: 2914905
  46. 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits a HeLa protein kinase that phosphorylates an RNA polymerase II-derived peptide.
    Biochem Biophys Res Commun. 1989 Mar 15;159(2):508-15 PMID: 2930526
  47. Inhibition of in vivo and in vitro transcription by monoclonal antibodies prepared against wheat germ RNA polymerase II that react with the heptapeptide repeat of eukaryotic RNA polymerase II.
    J Biol Chem. 1989 Jul 5;264(19):11511-20 PMID: 2472398
  48. Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry on thin sections.
    J Cell Biol. 1984 Jan;98(1):358-63 PMID: 6231300
  49. Transcription-dependent localization of U1 and U2 small nuclear ribonucleoproteins at major sites of gene activity in polytene chromosomes.
    J Mol Biol. 1984 Dec 25;180(4):911-26 PMID: 6241265
  50. Intranuclear localization of snRNP antigens.
    J Cell Biol. 1986 Jan;102(1):137-44 PMID: 2934400
  51. Casein kinase type II is involved in the inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole of specific RNA polymerase II transcription.
    J Biol Chem. 1986 Mar 5;261(7):3414-9 PMID: 3456346
  52. RNA contacts subunits IIo and IIc in HeLa RNA polymerase II transcription complexes.
    J Biol Chem. 1986 Oct 25;261(30):14226-31 PMID: 2429953
  53. Immunoelectron microscope visualization of nuclear ribonucleoprotein antigens within spread transcription complexes.
    J Cell Biol. 1986 Oct;103(4):1153-7 PMID: 2945824
  54. Messenger RNA synthesis in mammalian cells is catalyzed by the phosphorylated form of RNA polymerase II.
    J Biol Chem. 1987 Sep 15;262(26):12468-74 PMID: 3624268
  55. RNA polymerase II transcription cycles.
    Curr Opin Genet Dev. 1993 Apr;3(2):213-8 PMID: 8504246
  56. Fluorescent labeling of nascent RNA reveals transcription by RNA polymerase II in domains scattered throughout the nucleus.
    J Cell Biol. 1993 Jul;122(2):283-93 PMID: 8320255
  57. Nuclear organization of splicing small nuclear ribonucleoproteins in adenovirus-infected cells.
    J Virol. 1993 Oct;67(10):5792-802 PMID: 8371343
  58. RNA travel: tracks from DNA to cytoplasm.
    Cell. 1993 Nov 5;75(3):399-401 PMID: 7693353
  59. RNA polymerases IIA and IIO have distinct roles during transcription from the TATA-less murine dihydrofolate reductase promoter.
    J Biol Chem. 1993 Nov 25;268(33):25033-40 PMID: 8227067
  60. Locus-specific variation in phosphorylation state of RNA polymerase II in vivo: correlations with gene activity and transcript processing.
    Genes Dev. 1993 Dec;7(12A):2329-44 PMID: 8253380
  61. Tails of RNA polymerase II.
    Trends Biochem Sci. 1990 Oct;15(10):383-7 PMID: 2251729
  62. Identification of phosphorylation sites in the repetitive carboxyl-terminal domain of the mouse RNA polymerase II largest subunit.
    J Biol Chem. 1991 Feb 5;266(4):2290-6 PMID: 1899239
  63. Small nuclear ribonucleoproteins and heterogeneous nuclear ribonucleoproteins in the amphibian germinal vesicle: loops, spheres, and snurposomes.
    J Cell Biol. 1991 May;113(3):465-83 PMID: 1826687
  64. Spliceosomes and snurposomes.
    Science. 1991 Jun 14;252(5012):1499-500 PMID: 1828621
  65. RNA polymerase II.
    Annu Rev Biochem. 1991;60:689-715 PMID: 1883205
  66. Associations between distinct pre-mRNA splicing components and the cell nucleus.
    EMBO J. 1991 Nov;10(11):3467-81 PMID: 1833187
  67. A conserved family of nuclear phosphoproteins localized to sites of polymerase II transcription.
    J Cell Biol. 1991 Nov;115(3):587-96 PMID: 1717489
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.129.2.287
Published
1995-04-00
Pages
287-98
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2199908
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.129.2.287 PMC Full Text View journal details
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