Home LiteratureArticle Details
PMID: 8245117 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Nuclear organization of splicing snRNPs during differentiation of murine erythroleukemia cells in vitro.

The Journal of cell biology ·Vol. 123 ·No. 5 ·1993-12-00 ·Pages 1055-68

Antoniou M, Carmo-Fonseca M, Ferreira J, Lamond AI

Abstract

Murine erythroleukemia (MEL) cells are erythroid progenitors that can be induced to undergo terminal erythroid differentiation in culture. We have used MEL cells here as a model system to study the nuclear organization of splicing snRNPs during the physiological changes in gene expression which accompany differentiation. In uninduced MEL cells, snRNPs are widely distributed throughout the nucleoplasm and show an elevated concentration in coiled bodies. Within the first two days after induction of terminal erythroid differentiation, the pattern of gene expression changes, erythroid-specific transcription is activated and transcription of many other genes is repressed. During this early stage splicing snRNPs remain widely distributed through the nucleoplasm and continue to associate with coiled bodies. At later stages of differentiation (four to six days), when total transcription levels have greatly decreased, splicing snRNPs are redistributed. By six days postinduction snRNPs were concentrated in large clusters of interchromatin granules and no longer associated with coiled bodies. At the end-point of erythroid differentiation, just before enucleation, we observe a dramatic segregation of splicing snRNPs from the condensed chromatin. Analysis by EM shows that the snRNPs are packaged into a membrane-associated structure at the nuclear periphery which we term the "SCIM" domain (i.e., SnRNP Clusters Inside a Membrane).

MeSH Terms
Animals Cell Compartmentation Cell Nucleus/physiology,ultrastructure Chromatin/ultrastructure Erythroid Precursor Cells/physiology,ultrastructure Erythropoiesis/physiology Fluorescent Antibody Technique Gene Expression In Situ Hybridization Inclusion Bodies Mice Microscopy, Electron Nuclear Proteins/isolation & purification RNA Splicing Ribonucleoproteins Ribonucleoproteins, Small Nuclear/metabolism,ultrastructure Serine-Arginine Splicing Factors Tumor Cells, Cultured
Chemicals
Chromatin Nuclear Proteins Ribonucleoproteins Ribonucleoproteins, Small Nuclear SRSF2 protein, mouse Serine-Arginine Splicing Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Antoniou M
National Institute for Medical Research, The Ridgeway, London, United Kingdom.
Carmo-Fonseca M
Ferreira J
Lamond A I
References (47)
47 references, click to expand
  1. Identification of protein antigens associated with the nuclear matrix and with clusters of interchromatin granules in both interphase and mitotic cells.
    J Cell Sci. 1987 Mar;87 ( Pt 2):269-82 PMID: 3654787
  2. Biochemical characterization of U2 snRNP auxiliary factor: an essential pre-mRNA splicing factor with a novel intranuclear distribution.
    EMBO J. 1991 Jan;10(1):207-14 PMID: 1824937
  3. Globin RNA synthesis in vitro by isolated erythroleukemic cell nuclei: direct evidence for increased transcription during erythroid differentiation.
    Proc Natl Acad Sci U S A. 1977 Jun;74(6):2475-9 PMID: 267941
  4. Nucleoplasmic organization of small nuclear ribonucleoproteins in cultured human cells.
    J Cell Biol. 1993 May;121(4):715-27 PMID: 8491767
  5. Colocalization of U1 and U2 small nuclear RNPs by immunocytochemistry.
    Biol Cell. 1984;51(1):109-12 PMID: 6207877
  6. Terminal differentiation of murine erythroleukemia cells: physical stabilization of end-stage cells.
    J Cell Biol. 1982 May;93(2):390-4 PMID: 6954153
  7. Higher level organization of individual gene transcription and RNA splicing.
    Science. 1993 Feb 26;259(5099):1326-30 PMID: 8446901
  8. 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
  9. Structure of spliceosomal snRNPs and their role in pre-mRNA splicing.
    Biochim Biophys Acta. 1990 Nov 30;1087(3):265-92 PMID: 2147394
  10. Anti-RNP monoclonal antibodies derived from a mouse strain with lupus-like autoimmunity.
    J Immunol. 1982 Mar;128(3):1176-80 PMID: 6173424
  11. 5'-Terminal caps of snRNAs are reactive with antibodies specific for 2,2,7-trimethylguanosine in whole cells and nuclear matrices. Double-label immunofluorescent studies with anti-m3G antibodies and with anti-RNP and anti-Sm autoantibodies.
    Exp Cell Res. 1984 Oct;154(2):548-60 PMID: 6207038
  12. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide.
    Proc Natl Acad Sci U S A. 1971 Feb;68(2):378-82 PMID: 5277089
  13. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495-9 PMID: 316537
  14. Network antibodies identify nuclear lamin B as a physiological attachment site for peripherin intermediate filaments.
    Cell. 1991 Jan 11;64(1):109-21 PMID: 1986862
  15. A multisubunit factor, CstF, is required for polyadenylation of mammalian pre-mRNAs.
    Genes Dev. 1990 Dec;4(12A):2112-20 PMID: 1980119
  16. 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
  17. Nonhistone nuclear antigens reactive with autoantibodies. Immunofluorescence studies on distribution in synchronized cells.
    J Cell Biol. 1981 Dec;91(3 Pt 1):654-60 PMID: 7035465
  18. Loss of adhesion of murine erythroleukemia cells to fibronectin during erythroid differentiation.
    Science. 1984 Jun 1;224(4652):996-8 PMID: 6585955
  19. Definition of the minimal requirements within the human beta-globin gene and the dominant control region for high level expression.
    EMBO J. 1990 Jan;9(1):233-40 PMID: 2295312
  20. Mammalian nuclei contain foci which are highly enriched in components of the pre-mRNA splicing machinery.
    EMBO J. 1991 Jan;10(1):195-206 PMID: 1824936
  21. Splice site selection, rate of splicing, and alternative splicing on nascent transcripts.
    Genes Dev. 1988 Jun;2(6):754-65 PMID: 3138163
  22. 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
  23. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus.
    Nature. 1990 Feb 1;343(6257):437-41 PMID: 2137203
  24. A three-dimensional view of precursor messenger RNA metabolism within the mammalian nucleus.
    Science. 1993 Feb 26;259(5099):1330-5 PMID: 8446902
  25. 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
  26. Ultrastructural distribution of ribonucleoprotein complexes during mitosis. snRNP antigens are contained in mitotic granule clusters.
    Eur J Cell Biol. 1989 Dec;50(2):376-89 PMID: 2534076
  27. Erythropoietic differentiation in colonies of cells transformed by Friend virus.
    Proc Natl Acad Sci U S A. 1974 Jul;71(7):2668-70 PMID: 4527790
  28. The structure of mammalian small nuclear ribonucleoproteins. Identification of multiple protein components reactive with anti-(U1)ribonucleoprotein and anti-Sm autoantibodies.
    J Biol Chem. 1984 May 10;259(9):5907-14 PMID: 6232278
  29. Nascent pre-mRNA transcripts are associated with nuclear regions enriched in splicing factors.
    Genes Dev. 1991 Dec;5(12A):2288-302 PMID: 1748285
  30. beta-globin dominant control region interacts differently with distal and proximal promoter elements.
    Genes Dev. 1990 Jun;4(6):1007-13 PMID: 2384211
  31. 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
  32. In vivo detection of snRNP-rich organelles in the nuclei of mammalian cells.
    EMBO J. 1991 Jul;10(7):1863-73 PMID: 1710980
  33. A fibronectin matrix is required for differentiation of murine erythroleukemia cells into reticulocytes.
    J Cell Biol. 1987 Dec;105(6 Pt 2):3105-18 PMID: 2961771
  34. A new staining procedure for electron microscopical cytology.
    J Ultrastruct Res. 1969 May;27(3):250-65 PMID: 4181256
  35. Immunocytochemical identification of nuclear structures containing snRNPs in isolated rat liver cells.
    J Ultrastruct Res. 1984 May;87(2):180-9 PMID: 6544870
  36. Immunological and ultrastructural studies of the nuclear coiled body with autoimmune antibodies.
    Exp Cell Res. 1991 Jul;195(1):27-37 PMID: 2055273
  37. The coiled body.
    Trends Cell Biol. 1993 Jun;3(6):198-204 PMID: 14731494
  38. Cloning and intracellular localization of the U2 small nuclear ribonucleoprotein auxiliary factor small subunit.
    Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8769-73 PMID: 1388271
  39. Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry on thin sections.
    J Cell Biol. 1984 Jan;98(1):358-63 PMID: 6231300
  40. 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
  41. Changes in RNA and protein metabolism preceding onset of hemoglobin synthesis in cultured Friend leukemia cells.
    Dev Biol. 1976 Jan;48(1):118-31 PMID: 1245255
  42. Human autoantibody to a novel protein of the nuclear coiled body: immunological characterization and cDNA cloning of p80-coilin.
    J Exp Med. 1991 Jun 1;173(6):1407-19 PMID: 2033369
  43. Intranuclear distribution of U1 and U2 snRNAs visualized by high resolution in situ hybridization: revelation of a novel compartment containing U1 but not U2 snRNA in HeLa cells.
    Eur J Cell Biol. 1993 Apr;60(2):308-21 PMID: 8330629
  44. Highly localized tracks of specific transcripts within interphase nuclei visualized by in situ hybridization.
    Cell. 1989 May 5;57(3):493-502 PMID: 2541917
  45. Associations between distinct pre-mRNA splicing components and the cell nucleus.
    EMBO J. 1991 Nov;10(11):3467-81 PMID: 1833187
  46. Intranuclear localization of snRNP antigens.
    J Cell Biol. 1986 Jan;102(1):137-44 PMID: 2934400
  47. Xenopus tropicalis U6 snRNA genes transcribed by Pol III contain the upstream promoter elements used by Pol II dependent U snRNA genes.
    Nucleic Acids Res. 1987 Mar 25;15(6):2463-78 PMID: 3031599
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1993-12-00
Pages
1055-68
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2119874
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]