Home LiteratureArticle Details
PMID: 10329631 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inefficient processing impairs release of RNA from the site of transcription.

The EMBO journal ·Vol. 18 ·No. 10 ·1999-05-17 ·Pages 2855-66

Custódio N, Carmo-Fonseca M, Geraghty F, Pereira HS, Grosveld F, Antoniou M

Abstract

We describe here for the first time the site of retention within the nucleus of pre-mRNA processing mutants unable to be exported to the cytoplasm. Fluorescence in situ hybridization was used to detect transcripts from human beta-globin genes that are either normal or defective in splicing or 3' end formation. Nuclear transcripts of both wild-type and mutant RNAs are detected only as intranuclear foci that colocalize with the template gene locus. The kinetics of transcript release from the site of transcription was assessed by treatment of cells with the transcriptional inhibitors actinomycin D, alpha-amanitin and DRB. These drugs induce the rapid disappearance of nuclear foci corresponding to wild-type human beta-globin RNA. In contrast, pre-mRNA mutants defective in either splicing or 3' end formation and which fail to be transported to the cytoplasm, are retained at the site of transcription. Therefore, 3' end processing and splicing appear to be rate limiting for release of mRNA from the site of transcription.

MeSH Terms
Amanitins/pharmacology Cell Line Dactinomycin/pharmacology Dichlororibofuranosylbenzimidazole/pharmacology Globins/genetics Humans In Situ Hybridization, Fluorescence Kinetics Mutation RNA Precursors/metabolism RNA Processing, Post-Transcriptional/genetics RNA Splicing RNA, Messenger/metabolism Spliceosomes/genetics Transcription, Genetic
Chemicals
Amanitins RNA Precursors RNA, Messenger Dactinomycin Dichlororibofuranosylbenzimidazole Globins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Custódio N
Institute of Histology and Embryology, Faculty of Medicine, University of Lisbon, 1699 Lisbon codex, Portugal.
Carmo-Fonseca M
Geraghty F
Pereira H S
Grosveld F
Antoniou M
References (60)
60 references, click to expand
  1. Splicing precedes polyadenylation during Drosophila E74A transcription.
    Mol Cell Biol. 1990 Nov;10(11):6059-63 PMID: 1978245
  2. Monomethylated cap structures facilitate RNA export from the nucleus.
    Cell. 1990 Oct 5;63(1):109-18 PMID: 2208274
  3. Analysis of genes and chromosomes by nonisotopic in situ hybridization.
    Genet Anal Tech Appl. 1991 Feb;8(1):24-35 PMID: 2043382
  4. Mature mRNA 3' end formation stimulates RNA export from the nucleus.
    EMBO J. 1991 Nov;10(11):3513-22 PMID: 1833188
  5. Transport of RNA between nucleus and cytoplasm.
    Semin Cell Biol. 1992 Aug;3(4):279-88 PMID: 1384772
  6. Each hypersensitive site of the human beta-globin locus control region confers a different developmental pattern of expression on the globin genes.
    Genes Dev. 1993 Jan;7(1):106-13 PMID: 8422981
  7. Distribution of pre-mRNA splicing factors at sites of RNA polymerase II transcription.
    Genes Dev. 1997 May 1;11(9):1148-59 PMID: 9159396
  8. Pre-mRNA processing and the CTD of RNA polymerase II: the tail that wags the dog?
    Cell. 1997 May 16;89(4):491-4 PMID: 9160740
  9. Intergenic transcription and transinduction of the human beta-globin locus.
    Genes Dev. 1997 Oct 1;11(19):2494-509 PMID: 9334315
  10. Efficient 3'-end formation of human beta-globin mRNA in vivo requires sequences within the last intron but occurs independently of the splicing reaction.
    Nucleic Acids Res. 1998 Feb 1;26(3):721-9 PMID: 9443963
  11. Visualization of single RNA transcripts in situ.
    Science. 1998 Apr 24;280(5363):585-90 PMID: 9554849
  12. Chromatin interaction mechanism of transcriptional control in vivo.
    EMBO J. 1998 Oct 15;17(20):6020-7 PMID: 9774345
  13. Nuclear RNA export.
    Genes Dev. 1998 Nov 1;12(21):3303-19 PMID: 9808617
  14. Action of nucleic acids on the inhibition of growth by actinomycin of Neurospora crassa.
    Nature. 1960 Jul 2;187:60-1 PMID: 14408651
  15. Basis of actinomycin action. I. DNA binding and inhibition of RNA-polymerase synthetic reactions by actinomycin.
    Proc Natl Acad Sci U S A. 1962 Dec 15;48:2094-101 PMID: 13948669
  16. Higher level organization of individual gene transcription and RNA splicing.
    Science. 1993 Feb 26;259(5099):1326-30 PMID: 8446901
  17. The basic domain of Rev from human immunodeficiency virus type 1 specifically blocks the entry of U4/U6.U5 small nuclear ribonucleoprotein in spliceosome assembly.
    J Virol. 1993 Aug;67(8):4769-76 PMID: 8331728
  18. Nuclear organization of splicing snRNPs during differentiation of murine erythroleukemia cells in vitro.
    J Cell Biol. 1993 Dec;123(5):1055-68 PMID: 8245117
  19. Splicing of Balbiani ring 1 gene pre-mRNA occurs simultaneously with transcription.
    Cell. 1994 Jan 14;76(1):183-92 PMID: 8287477
  20. Nuclear export of different classes of RNA is mediated by specific factors.
    J Cell Biol. 1994 Mar;124(5):627-35 PMID: 7509815
  21. Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries.
    Chromosome Res. 1993 Jul;1(2):93-106 PMID: 8143096
  22. mRNA nuclear export.
    Curr Opin Genet Dev. 1994 Apr;4(2):305-9 PMID: 8032209
  23. Association of nuclear matrix antigens with exon-containing splicing complexes.
    J Cell Biol. 1994 Nov;127(3):593-607 PMID: 7962048
  24. Localization of pre-mRNA splicing in mammalian nuclei.
    Nature. 1994 Dec 22-29;372(6508):809-12 PMID: 7997273
  25. Dynamic RNA-RNA interactions in the spliceosome.
    Annu Rev Genet. 1994;28:1-26 PMID: 7534458
  26. The human dystrophin gene requires 16 hours to be transcribed and is cotranscriptionally spliced.
    Nat Genet. 1995 Feb;9(2):184-90 PMID: 7719347
  27. Rearrangement of snRNA structure during assembly and function of the spliceosome.
    Prog Nucleic Acid Res Mol Biol. 1995;50:131-59 PMID: 7754032
  28. Regulation of transcriptional elongation by RNA polymerase II.
    Curr Opin Genet Dev. 1995 Apr;5(2):210-6 PMID: 7613091
  29. Transcription complex stability and chromatin dynamics in vivo.
    Nature. 1995 Sep 21;377(6546):209-13 PMID: 7675106
  30. A pre-mRNA-binding protein accompanies the RNA from the gene through the nuclear pores and into polysomes.
    Cell. 1996 Jan 26;84(2):253-64 PMID: 8565071
  31. A nuclear cap-binding complex binds Balbiani ring pre-mRNA cotranscriptionally and accompanies the ribonucleoprotein particle during nuclear export.
    J Cell Biol. 1996 Apr;133(1):5-14 PMID: 8601613
  32. Intron-dependent recruitment of pre-mRNA splicing factors to sites of transcription.
    J Cell Biol. 1996 May;133(4):719-32 PMID: 8666659
  33. Demonstration of a dynamic, transcription-dependent organization of pre-mRNA splicing factors in polytene nuclei.
    J Cell Biol. 1996 Jun;133(5):929-41 PMID: 8655585
  34. The C-terminal domain of the largest subunit of RNA polymerase II interacts with a novel set of serine/arginine-rich proteins.
    Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):6975-80 PMID: 8692929
  35. A hyperphosphorylated form of the large subunit of RNA polymerase II is associated with splicing complexes and the nuclear matrix.
    Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8253-7 PMID: 8710856
  36. The intranuclear site of excision of each intron in Balbiani ring 3 pre-mRNA is influenced by the time remaining to transcription termination and different excision efficiencies for the various introns.
    RNA. 1996 Jul;2(7):641-51 PMID: 8756407
  37. In vivo degradation of RNA polymerase II largest subunit triggered by alpha-amanitin.
    Nucleic Acids Res. 1996 Aug 1;24(15):2924-9 PMID: 8760875
  38. Heterochromatin effects on the frequency and duration of LCR-mediated gene transcription.
    Cell. 1996 Oct 4;87(1):105-14 PMID: 8858153
  39. Localization of HIV-1 RNA in mammalian nuclei.
    J Cell Biol. 1996 Oct;135(1):9-18 PMID: 8858159
  40. The nuclear matrix protein p255 is a highly phosphorylated form of RNA polymerase II largest subunit which associates with spliceosomes.
    Nucleic Acids Res. 1996 Dec 1;24(23):4649-52 PMID: 8972849
  41. The C-terminal domain of RNA polymerase II couples mRNA processing to transcription.
    Nature. 1997 Jan 23;385(6614):357-61 PMID: 9002523
  42. A functional interaction between the carboxy-terminal domain of RNA polymerase II and pre-mRNA splicing.
    J Cell Biol. 1997 Jan 13;136(1):5-18 PMID: 9008699
  43. RNA transport.
    Annu Rev Neurosci. 1997;20:269-301 PMID: 9056715
  44. Nucleocytoplasmic transport: signals, mechanisms and regulation.
    Nature. 1997 Apr 24;386(6627):779-87 PMID: 9126736
  45. RNA movement between the nucleus and the cytoplasm.
    Curr Opin Genet Dev. 1997 Apr;7(2):212-9 PMID: 9115427
  46. Alpha-amanitin: a specific inhibitor of one of two DNA-pendent RNA polymerase activities from calf thymus.
    Biochem Biophys Res Commun. 1970 Jan 6;38(1):165-71 PMID: 4907405
  47. Specific inhibition of nuclear RNA polymerase II by alpha-amanitin.
    Science. 1970 Oct 23;170(3956):447-9 PMID: 4918258
  48. Polyadenylic acid sequences: role in conversion of nuclear RNA into messenger RNA.
    Science. 1971 Oct 29;174(4008):507-10 PMID: 5110429
  49. Animal DNA-dependent RNA polymerases. 11. Mechanism of the inhibition of RNA polymerases B by amatoxins.
    Biochim Biophys Acta. 1974 Jun 27;353(2):160-84 PMID: 4601749
  50. Photoreactivation of amanitin-inhibited RNA polymerase II.
    J Biol Chem. 1982 Feb 25;257(4):1577-8 PMID: 7056732
  51. 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
  52. A mutational analysis of spliceosome assembly: evidence for splice site collaboration during spliceosome formation.
    Genes Dev. 1987 Aug;1(6):532-43 PMID: 2824284
  53. Splice site selection, rate of splicing, and alternative splicing on nascent transcripts.
    Genes Dev. 1988 Jun;2(6):754-65 PMID: 3138163
  54. A dominant control region from the human beta-globin locus conferring integration site-independent gene expression.
    Nature. 1989 Mar 23;338(6213):352-5 PMID: 2922063
  55. The beta-globin dominant control region activates homologous and heterologous promoters in a tissue-specific manner.
    Cell. 1989 Mar 24;56(6):969-77 PMID: 2924354
  56. Some cis- and trans-acting mutants for splicing target pre-mRNA to the cytoplasm.
    Cell. 1989 May 19;57(4):573-83 PMID: 2655924
  57. Evidence that a nuclear matrix protein participates in premessenger RNA splicing.
    Exp Cell Res. 1989 Jun;182(2):521-33 PMID: 2721591
  58. Regulation by HIV Rev depends upon recognition of splice sites.
    Cell. 1989 Dec 1;59(5):789-95 PMID: 2686839
  59. 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
  60. 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
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/18.10.2855
Published
1999-05-17
Pages
2855-66
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171366
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/18.10.2855 PMC Full Text View journal details
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: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2