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PMID: 6203096 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The 5'-terminal sequence of U1 RNA complementary to the consensus 5' splice site of hnRNA is single-stranded in intact U1 snRNP particles.

Nucleic acids research ·Vol. 12 ·No. 10 ·1984-05-25 ·Pages 4111-26

Rinke J, Appel B, Blöcker H, Frank R, Lührmann R

Abstract

The 5'-terminal region of U1 snRNA is highly complementary to the consensus exon-intron regions of hnRNA and it has been suggested that U1 snRNP might play a role in the splicing of the pre-mRNA by intermolecular base-pairing between these regions. Here the secondary structure of the 5' terminus of U1 RNA in the isolated native U1 snRNP particle has been investigated by site-directed enzymatic cleavage of the RNA. Individual oligodeoxynucleotides complementary to various sequences within the first 15 nucleotides of the 5' terminus of U1 RNA have been tested for their ability to form stable DNA X RNA hybrids, with subsequent cleavage of the U1 RNA by RNase H. Our results show unequivocally that the 9 nucleotides at the 5' terminus which are complementary to a consensus 5' splice site are indeed single-stranded in the intact U1 snRNP particle, and are not protected by snRNP proteins. However, they also indicate that the U1 sequence complementary to an intron's consensus 3' end is not readily available for intermolecular base-pairing, either in the intact U1 snRNP particle or in the deproteinized U1 RNA molecule. Therefore our data favour the possibility that U1 snRNP plays a role only in the recognition of a 5' splice site of hnRNA, rather than being involved in the alignment of both ends of an intron for splicing.

MeSH Terms
Autoantibodies Base Composition Base Sequence Connective Tissue Diseases/immunology Endoribonucleases HeLa Cells/physiology Humans Nucleic Acid Conformation Nucleic Acid Hybridization RNA/genetics RNA Splicing RNA, Small Nuclear Ribonuclease H Ribonucleoproteins/genetics,isolation & purification Ribonucleoproteins, Small Nuclear
Chemicals
Autoantibodies RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear RNA Endoribonucleases Ribonuclease H
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Rinke J
Appel B
Blöcker H
Frank R
Lührmann R
References (32)
32 references, click to expand
  1. Stimultaneous purification of Escherichia coli termination factor rho, RNAase III and RNAase H.
    Eur J Biochem. 1975 Feb 21;51(2):369-76 PMID: 1097242
  2. A catalogue of splice junction sequences.
    Nucleic Acids Res. 1982 Jan 22;10(2):459-72 PMID: 7063411
  3. The presence of small molecular weight RNAs in nuclear ribonucleoprotein particles carrying HnRNA.
    FEBS Lett. 1977 Jan 15;73(1):80-4 PMID: 838050
  4. Circular dichroism of ribonucleoprotein complexes from rat liver nuclei.
    Biochem Biophys Res Commun. 1977 Jun 20;76(4):1130-7 PMID: 901464
  5. Mapping adenines, guanines, and pyrimidines in RNA.
    Nucleic Acids Res. 1977 Aug;4(8):2527-38 PMID: 409999
  6. Accessibility of U1 RNA to base pairing with a single-stranded DNA fragment mimicking the intron extremities at the splice junction.
    Nucleic Acids Res. 1982 Feb 25;10(4):1193-201 PMID: 6175953
  7. Small nuclear ribonucleoproteins of Drosophila: identification of U1 RNA-associated proteins and their behavior during heat shock.
    Mol Cell Biol. 1982 Aug;2(8):914-20 PMID: 6182453
  8. Ultrafast sequencing of oligodeoxyribonucleotides by FAB-mass spectrometry.
    Nucleic Acids Res. 1982 Aug 11;10(15):4671-8 PMID: 7133994
  9. Recent developments in the chemical synthesis of polynucleotides.
    Nucleic Acids Res. 1982 Nov 11;10(21):6553-70 PMID: 6757865
  10. Isolation and characterization of rabbit anti-m3 2,2,7G antibodies.
    Nucleic Acids Res. 1982 Nov 25;10(22):7103-13 PMID: 7155893
  11. 5'-terminal caps of snRNAs are accessible for reaction with 2,2,7-trimethylguanosine-specific antibody in intact snRNPs.
    J Biol Chem. 1983 Mar 10;258(5):2745-7 PMID: 6186660
  12. Structural organization of ribonucleoproteins containing small nuclear RNAs from HeLa cells. Proteins interact closely with a similar structural domain of U1, U2, U4 and U5 small nuclear RNAs.
    J Mol Biol. 1982 Dec 15;162(3):623-43 PMID: 6187926
  13. Purification of snRNPs U1, U2, U4, U5 and U6 with 2,2,7-trimethylguanosine-specific antibody and definition of their constituent proteins reacting with anti-Sm and anti-(U1)RNP antisera.
    EMBO J. 1983;2(7):1129-35 PMID: 6194992
  14. U2 RNA shares a structural domain with U1, U4, and U5 RNAs.
    EMBO J. 1982;1(10):1259-65 PMID: 6202507
  15. High-pressure liquid chromatography in polynucleotide synthesis.
    Biochemistry. 1978 Apr 4;17(7):1257-67 PMID: 656387
  16. Addressed fragmentation of RNA Molecules.
    FEBS Lett. 1979 Jul 1;103(1):197-9 PMID: 467648
  17. An evaluation of small nuclear RNA in hnRNP.
    FEBS Lett. 1979 Aug 1;104(1):176-82 PMID: 477978
  18. Site specific enzymatic cleavage of RNA.
    Nucleic Acids Res. 1979 Sep 11;7(1):179-92 PMID: 386279
  19. Are snRNPs involved in splicing?
    Nature. 1980 Jan 10;283(5743):220-4 PMID: 7350545
  20. 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
  21. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  22. A mechanism for RNA splicing.
    Proc Natl Acad Sci U S A. 1980 Apr;77(4):1877-9 PMID: 6246511
  23. Nucleotide sequences of nuclear U1A RNAs from chicken, rat and man.
    Nucleic Acids Res. 1980 Sep 25;8(18):4143-54 PMID: 6159587
  24. Site-specific cleavage by T1 RNase of U-1 RNA in u-1 ribonucleoprotein particles.
    Proc Natl Acad Sci U S A. 1981 Mar;78(3):1562-6 PMID: 6165013
  25. Comparison of RNase T1 fingerprints of U1, U2, and U3 small nuclear RNA's of HeLa cells, human normal fibroblasts, and Novikoff hepatoma cells.
    Cancer Res. 1981 Jun;41(6):2215-20 PMID: 6165460
  26. Sequence of U1 RNA from Drosophila melanogaster: implications for U1 secondary structure and possible involvement in splicing.
    Nucleic Acids Res. 1981 Dec 11;9(23):6351-68 PMID: 6172778
  27. Localisation of a series of intra-RNA cross-links in the secondary and tertiary structure of 23S RNA, induced by ultraviolet irradiation of Escherichia coli 50S ribosomal subunits.
    Nucleic Acids Res. 1983 Mar 25;11(6):1687-706 PMID: 6340065
  28. The U1 small nuclear RNA-protein complex selectively binds a 5' splice site in vitro.
    Cell. 1983 Jun;33(2):509-18 PMID: 6190573
  29. Splicing of messenger RNA precursors is inhibited by antisera to small nuclear ribonucleoprotein.
    Cell. 1983 Nov;35(1):101-7 PMID: 6194895
  30. Splicing of in vitro synthesized messenger RNA precursors in HeLa cell extracts.
    Cell. 1983 Nov;35(1):89-99 PMID: 6194902
  31. Base-pairing interactions between small nuclear RNAs and nuclear RNA precursors as revealed by psoralen cross-linking in vivo.
    Cell. 1981 Nov;26(3 Pt 1):363-70 PMID: 6173132
  32. Nuclear ribonucleoprotein complexes containing U1 and U2 RNA.
    Biochemistry. 1975 Oct 7;14(20):4380-5 PMID: 169894
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1984-05-25
Pages
4111-26
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318820
Subset
IM
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