Home LiteratureArticle Details
PMID: 2461296 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Antisense RNA inhibits splicing of pre-mRNA in vitro.

The EMBO journal ·Vol. 7 ·No. 8 ·1988-08-00 ·Pages 2523-32

Munroe SH

Abstract

Antisense RNAs complementary to human beta-globin pre-mRNA or to a chimeric globin/adenovirus E2a pre-mRNA specifically and efficiently inhibit pre-mRNA splicing in vitro. The level of inhibition depends on the length, position and concentration of the antisense RNA relative to the pre-mRNA substrate. Antisense RNAs complementary to sequences greater than 80 nucleotides downstream of the globin 3' splice site inhibit at least as efficiently as those extending across the splice sites. Thus splicing is sensitive to perturbations involving exon sequences some distance from the splice sites. Inhibition is mediated by factors which affect the annealing of antisense and substrate RNAs. Direct analysis of RNA duplex formation demonstrates the presence of an activity in HeLa cell nuclear extract which promotes the rapid annealing of complementary RNAs in an ATP-independent manner. Both annealing and inhibition are greatly reduced when antisense RNA is added to the splicing reaction greater than or equal to 5 min after substrate. This result may reflect a transition between an open structure, in which annealing of antisense RNA with pre-mRNA is facilitated, and a closed complex in which pre-mRNA is sequestered at an early stage of spliceosome assembly.

MeSH Terms
Adenoviridae/genetics Base Sequence Exons Globins/genetics HeLa Cells Humans Molecular Sequence Data Nucleic Acid Hybridization RNA/pharmacology RNA Precursors/genetics RNA Splicing/drug effects RNA, Antisense RNA, Messenger/genetics Transcription, Genetic
Chemicals
RNA Precursors RNA, Antisense RNA, Messenger RNA Globins
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Munroe S H
Cold Spring Harbor Laboratory, NY 11724.
References (43)
43 references, click to expand
  1. In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.
    Proc Natl Acad Sci U S A. 1984 Aug;81(15):4707-11 PMID: 6589618
  2. A minimal intron length but no specific internal sequence is required for splicing the large rabbit beta-globin intron.
    Cell. 1984 Jul;37(3):915-25 PMID: 6204770
  3. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  4. Secondary structure of splice sites in adenovirus mRNA precursors.
    Nucleic Acids Res. 1984 Nov 26;12(22):8437-56 PMID: 6095200
  5. Role of intron-contained sequences in formation of moloney murine leukemia virus env mRNA.
    Mol Cell Biol. 1984 Nov;4(11):2289-97 PMID: 6096692
  6. Injected anti-sense RNAs specifically block messenger RNA translation in vivo.
    Proc Natl Acad Sci U S A. 1985 Jan;82(1):144-8 PMID: 3855537
  7. Trans splicing of mRNA precursors in vitro.
    Cell. 1985 Aug;42(1):165-71 PMID: 3848348
  8. A multicomponent complex is involved in the splicing of messenger RNA precursors.
    Cell. 1985 Aug;42(1):345-53 PMID: 3160482
  9. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences.
    Cell. 1985 Aug;42(1):355-67 PMID: 3160483
  10. Exon mutations that affect the choice of splice sites used in processing the SV40 late transcripts.
    Nucleic Acids Res. 1985 Aug 12;13(15):5591-609 PMID: 2994004
  11. Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro.
    Cell. 1985 Oct;42(3):725-36 PMID: 2996774
  12. Specific and stable intron-factor interactions are established early during in vitro pre-mRNA splicing.
    Cell. 1985 Nov;43(1):131-42 PMID: 3000599
  13. Alternative splicing caused by RNA secondary structure.
    Cell. 1985 Dec;43(3 Pt 2):667-76 PMID: 4075405
  14. Inhibition of splicing but not cleavage at the 5' splice site by truncating human beta-globin pre-mRNA.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):927-31 PMID: 2937057
  15. Heterogeneous nuclear ribonucleoproteins: role in RNA splicing.
    Science. 1986 Mar 28;231(4745):1534-9 PMID: 3952495
  16. Splicing of the E2A premessenger RNA of adenovirus serotype 2. Multiple pathways in spite of excision of the entire large intron.
    J Mol Biol. 1986 Feb 5;187(3):379-97 PMID: 3009832
  17. Purification and domain structure of core hnRNP proteins A1 and A2 and their relationship to single-stranded DNA-binding proteins.
    J Biol Chem. 1986 Aug 25;261(24):11266-73 PMID: 3733753
  18. Antibodies to hnRNP core proteins inhibit in vitro splicing of human beta-globin pre-mRNA.
    Nucleic Acids Res. 1986 Jul 11;14(13):5241-54 PMID: 3737400
  19. Single-stranded DNA binding proteins required for DNA replication.
    Annu Rev Biochem. 1986;55:103-36 PMID: 3527040
  20. Splicing of messenger RNA precursors.
    Annu Rev Biochem. 1986;55:1119-50 PMID: 2943217
  21. The role of antisense RNA in gene regulation.
    Annu Rev Biochem. 1986;55:569-97 PMID: 2427015
  22. A role for exon sequences and splice-site proximity in splice-site selection.
    Cell. 1986 Aug 29;46(5):681-90 PMID: 2427200
  23. Sequence requirements for splicing of higher eukaryotic nuclear pre-mRNA.
    Cell. 1986 Nov 21;47(4):555-65 PMID: 3779836
  24. The role of nucleotide sequences in splice site selection in eukaryotic pre-messenger RNA.
    Nature. 1986 Nov 20-26;324(6094):280-2 PMID: 2946960
  25. Splice site consensus sequences are preferentially accessible to nucleases in isolated adenovirus RNA.
    Nucleic Acids Res. 1986 Nov 11;14(21):8447-65 PMID: 3024107
  26. Pre-mRNA splicing.
    Annu Rev Genet. 1986;20:671-708 PMID: 2880558
  27. Multiple interactions between the splicing substrate and small nuclear ribonucleoproteins in spliceosomes.
    Mol Cell Biol. 1987 Jan;7(1):281-93 PMID: 2951586
  28. Primary structure of human nuclear ribonucleoprotein particle C proteins: conservation of sequence and domain structures in heterogeneous nuclear RNA, mRNA, and pre-rRNA-binding proteins.
    Mol Cell Biol. 1987 May;7(5):1731-9 PMID: 3110598
  29. Minimal exon sequence requirements for efficient in vitro splicing of mono-intronic nuclear pre-mRNA.
    J Biol Chem. 1987 Aug 15;262(23):11284-91 PMID: 3649340
  30. Amount of RNA secondary structure required to induce an alternative splice.
    Mol Cell Biol. 1987 Sep;7(9):3194-8 PMID: 3670310
  31. 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
  32. Purification and RNA binding properties of a C-type hnRNP protein from HeLa cells.
    J Biol Chem. 1987 Dec 15;262(35):17126-37 PMID: 2445755
  33. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA.
    Nature. 1970 Sep 26;227(5265):1313-8 PMID: 5455134
  34. Catalysis of DNA reassociation by the Escherichia coli DNA binding protein: A polyamine-dependent reaction.
    J Mol Biol. 1977 Sep 25;115(3):441-54 PMID: 22760
  35. Processing and expression of early SV40 mRNA: a role for RNA conformation in splicing.
    Cell. 1979 Sep;18(1):85-92 PMID: 228860
  36. A RNA helix-destabilizing protein is a major component of Artemia salina nuclear ribonucleoproteins.
    Proc Natl Acad Sci U S A. 1981 May;78(5):2888-92 PMID: 6265922
  37. The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction.
    Nucleic Acids Res. 1987 Nov 25;15(22):9279-98 PMID: 3317279
  38. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  39. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene.
    Cell. 1983 Oct;34(3):865-79 PMID: 6313211
  40. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis.
    Cell. 1984 Apr;36(4):1007-15 PMID: 6323013
  41. Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro.
    Cell. 1984 Apr;36(4):993-1005 PMID: 6323033
  42. A large inverted repeat sequence overlaps two acceptor splice sites in adenovirus.
    Nucleic Acids Res. 1983 Dec 20;11(24):8891-900 PMID: 6672774
  43. Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro.
    Cell. 1984 Aug;38(1):317-31 PMID: 6088074
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1988-08-00
Pages
2523-32
Language
English
Region
England
NLM ID
8208664
PMCID
PMC457123
Subset
IM
Grants
NCI NIH HHS · CA 13106 · United States
NIGMS NIH HHS · GM 12126 · United States
External Links
PubMed source 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