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PMID: 11598018 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Antagonistic effects of T-Ag and VP16 reveal a role for RNA pol II elongation on alternative splicing.

The EMBO journal ·Vol. 20 ·No. 20 ·2001-10-15 ·Pages 5759-68

Kadener S, Cramer P, Nogués G, Cazalla D, de la Mata M, Fededa JP, Werbajh SE, Srebrow A, Kornblihtt AR

Abstract

Here we investigate the promoter control of alternative splicing by studying two transcriptional activators on templates under replicating conditions. SV40 large T-antigen (T-Ag) activates template replication only 2-fold but transcription 25-fold. T-Ag-mediated replication, reported to inhibit RNA polymerase II elongation, provokes a 10- to 30-fold increase in the inclusion of the fibronectin EDI exon into mature mRNA. The T-Ag effect is exon specific, occurs in cis and depends strictly on DNA replication and not on cell transformation. VP16, an activator of transcriptional initiation and elongation, has a similar effect on transcription but the opposite effect on splicing: EDI inclusion is inhibited by 35-fold. VP16 completely reverts the T-Ag effect, but a VP16 mutant with reduced elongation ability provokes only partial reversion. Both T-Ag and VP16 promote conspicuous co-localization of mRNA with nuclear speckles that contain the SR protein SF2/ASF, a positive regulator of EDI inclusion. Therefore, we conclude that co-localization of transcripts and speckles is not sufficient to stimulate EDI inclusion.

MeSH Terms
Alternative Splicing Animals Antigens, Polyomavirus Transforming/physiology COS Cells Carcinoma, Hepatocellular/genetics Chlorocebus aethiops DNA Replication DNA, Recombinant/genetics,metabolism Exons/genetics Fibronectins/genetics,metabolism Herpes Simplex Virus Protein Vmw65/physiology Humans In Situ Hybridization Liver Neoplasms/genetics Promoter Regions, Genetic/genetics RNA Polymerase II/metabolism RNA, Messenger/biosynthesis Recombinant Fusion Proteins/physiology Reverse Transcriptase Polymerase Chain Reaction Simian virus 40 Templates, Genetic Transcription, Genetic Transfection
Chemicals
Antigens, Polyomavirus Transforming DNA, Recombinant Fibronectins Herpes Simplex Virus Protein Vmw65 RNA, Messenger Recombinant Fusion Proteins RNA Polymerase II
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Kadener S
Laboratorio de Fisiología y Biología Molecular, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellón 2, (C1428EHA) Buenos Aires, Argentina.
Cramer P
Nogués G
Cazalla D
de la Mata M
Fededa J P
Werbajh S E
Srebrow A
Kornblihtt A R
References (44)
44 references, click to expand
  1. Chromatin modification by DNA tracking.
    Proc Natl Acad Sci U S A. 1999 Nov 23;96(24):13634-7 PMID: 10570124
  2. Structure and function of simian virus 40 large tumor antigen.
    Annu Rev Biochem. 1992;61:55-85 PMID: 1323237
  3. High mobility of proteins in the mammalian cell nucleus.
    Nature. 2000 Apr 6;404(6778):604-9 PMID: 10766243
  4. Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function.
    J Cell Sci. 2000 Jun;113 ( Pt 11):1841-9 PMID: 10806095
  5. Distribution of acetylated histones resulting from Gal4-VP16 recruitment of SAGA and NuA4 complexes.
    EMBO J. 2000 Jun 1;19(11):2629-40 PMID: 10835360
  6. Connecting transcription to messenger RNA processing.
    Trends Biochem Sci. 2000 Jun;25(6):290-3 PMID: 10838569
  7. RNA polymerase II and the integration of nuclear events.
    Genes Dev. 2000 Jun 15;14(12):1415-29 PMID: 10859161
  8. Drosophila Dscam is an axon guidance receptor exhibiting extraordinary molecular diversity.
    Cell. 2000 Jun 9;101(6):671-84 PMID: 10892653
  9. Replication but not transcription of simian virus 40 DNA is dependent on nuclear domain 10.
    J Virol. 2000 Oct;74(20):9694-700 PMID: 11000241
  10. Protein diversity from alternative splicing: a challenge for bioinformatics and post-genome biology.
    Cell. 2000 Oct 27;103(3):367-70 PMID: 11081623
  11. Coordination between transcription and pre-mRNA processing.
    FEBS Lett. 2001 Jun 8;498(2-3):179-82 PMID: 11412852
  12. A splicing enhancer in the human fibronectin alternate ED1 exon interacts with SR proteins and stimulates U2 snRNP binding.
    Genes Dev. 1993 Dec;7(12A):2405-17 PMID: 8253386
  13. A novel bipartite splicing enhancer modulates the differential processing of the human fibronectin EDA exon.
    Nucleic Acids Res. 1994 Mar 25;22(6):1018-22 PMID: 8152907
  14. Transcriptional elongation by RNA polymerase II is stimulated by transactivators.
    Cell. 1994 Jun 3;77(5):749-59 PMID: 8205623
  15. Effects of the simian virus 40 origin of replication on transcription from the human immunodeficiency virus type 1 promoter.
    J Virol. 1995 Feb;69(2):1296-301 PMID: 7815509
  16. Regulation of fibronectin expression in rat regenerating liver.
    Nucleic Acids Res. 1995 Jan 25;23(2):238-43 PMID: 7862527
  17. Promoter-proximal pausing of RNA polymerase II defines a general rate-limiting step after transcription initiation.
    Genes Dev. 1995 Mar 1;9(5):559-72 PMID: 7698646
  18. Activation domains of transcription factors mediate replication dependent transcription from a minimal HIV-1 promoter.
    Nucleic Acids Res. 1996 Feb 15;24(4):549-57 PMID: 8604293
  19. Three functional classes of transcriptional activation domain.
    Mol Cell Biol. 1996 May;16(5):2044-55 PMID: 8628270
  20. The fibronectin gene as a model for splicing and transcription studies.
    FASEB J. 1996 Feb;10(2):248-57 PMID: 8641558
  21. The C-terminal domain of RNA polymerase II couples mRNA processing to transcription.
    Nature. 1997 Jan 23;385(6614):357-61 PMID: 9002523
  22. The dynamics of a pre-mRNA splicing factor in living cells.
    Nature. 1997 May 29;387(6632):523-7 PMID: 9168118
  23. HRS/SRp40-mediated inclusion of the fibronectin EIIIB exon, a possible cause of increased EIIIB expression in proliferating liver.
    Mol Cell Biol. 1997 Jul;17(7):4096-104 PMID: 9199345
  24. Functional association between promoter structure and transcript alternative splicing.
    Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11456-60 PMID: 9326631
  25. A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm.
    Genes Dev. 1998 Jan 1;12(1):55-66 PMID: 9420331
  26. Alternative splicing of the fibronectin EIIIB exon depends on specific TGCATG repeats.
    Mol Cell Biol. 1998 Jul;18(7):3900-6 PMID: 9632774
  27. Long-distance transcriptional enhancement by the histone acetyltransferase PCAF.
    Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13501-6 PMID: 9811829
  28. Co-transcriptional commitment to alternative splice site selection.
    Nucleic Acids Res. 1998 Dec 15;26(24):5568-72 PMID: 9837984
  29. Processing of endogenous pre-mRNAs in association with SC-35 domains is gene specific.
    J Cell Biol. 1999 Feb 22;144(4):617-29 PMID: 10037785
  30. RNA polymerase II targets pre-mRNA splicing factors to transcription sites in vivo.
    Mol Cell. 1999 Jun;3(6):697-705 PMID: 10394358
  31. Coupling RNA polymerase II transcription with pre-mRNA processing.
    Curr Opin Cell Biol. 1999 Jun;11(3):347-51 PMID: 10395561
  32. Coupling of transcription with alternative splicing: RNA pol II promoters modulate SF2/ASF and 9G8 effects on an exonic splicing enhancer.
    Mol Cell. 1999 Aug;4(2):251-8 PMID: 10488340
  33. Selective extraction of polyoma DNA from infected mouse cell cultures.
    J Mol Biol. 1967 Jun 14;26(2):365-9 PMID: 4291934
  34. Human fibronectin: molecular cloning evidence for two mRNA species differing by an internal segment coding for a structural domain.
    EMBO J. 1984 Jan;3(1):221-6 PMID: 6200322
  35. Assembly of transfected DNA into chromatin: structural changes in the origin-promoter-enhancer region upon replication.
    EMBO J. 1984 Jun;3(6):1243-53 PMID: 6086311
  36. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  37. A role for exon sequences in alternative splicing of the human fibronectin gene.
    Nucleic Acids Res. 1987 Oct 12;15(19):7725-33 PMID: 3671064
  38. Identification of a third region of cell-specific alternative splicing in human fibronectin mRNA.
    Proc Natl Acad Sci U S A. 1987 Oct;84(20):7179-82 PMID: 3478690
  39. Sequence analysis and in vivo expression show that alternative splicing of ED-B and ED-A regions of the human fibronectin gene are independent events.
    Nucleic Acids Res. 1988 Apr 25;16(8):3545-57 PMID: 3375063
  40. Effects of RNA secondary structure on alternative splicing of pre-mRNA: is folding limited to a region behind the transcribing RNA polymerase?
    Cell. 1988 Jul 29;54(3):393-401 PMID: 2840206
  41. Nucleosome assembly in mammalian cell extracts before and after DNA replication.
    EMBO J. 1990 Sep;9(9):2911-22 PMID: 2167837
  42. The ability of simian virus 40 large T antigen to immortalize primary mouse embryo fibroblasts cosegregates with its ability to bind to p53.
    J Virol. 1991 Dec;65(12):6872-80 PMID: 1658380
  43. Multiple SP1 binding sites confer enhancer-independent, replication-activated transcription of HIV-1 and globin gene promoters.
    New Biol. 1992 Apr;4(4):369-81 PMID: 1622932
  44. ISIS, the intron information system, reveals the high frequency of alternative splicing in the human genome.
    Nat Genet. 2000 Apr;24(4):340-1 PMID: 10742092
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.20.5759
Published
2001-10-15
Pages
5759-68
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125675
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.20.5759 PMC Full Text View journal details
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