Home LiteratureArticle Details
PMID: 3141901 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 and U5 snRNPs.

Nucleic acids research ·Vol. 16 ·No. 20 ·1988-10-25 ·Pages 9415-29

Krainer AR

Abstract

The four major nucleoplasmic small nuclear ribonucleoprotein particles U1, U2, U4/U6 and U5 can be extensively purified from HeLa cells by immunoaffinity chromatography using a monoclonal anti-trimethylguanosine antibody. The snRNP particles in active splicing extracts are selectively bound to the immunoaffinity matrix, and are then gently eluted by competition with an excess of free nucleoside. Biochemical complementation studies show that the purified snRNPs are active in pre-mRNA splicing, but only in the presence of additional non-snRNP protein factors. All the RNPs that are necessary for splicing can be purified in this manner. The active snRNPs are characterized with respect to their polypeptide composition, and shown to be distinct from several other activities implicated in splicing.

MeSH Terms
Animals Carrier Proteins/isolation & purification Chromatography, Affinity Genetic Complementation Test HeLa Cells Humans Mice Mice, Inbred BALB C Peptides/metabolism RNA Splicing RNA, Messenger/metabolism RNA, Small Nuclear/isolation & purification,metabolism,physiology RNA-Binding Proteins
Chemicals
Carrier Proteins Peptides RNA, Messenger RNA, Small Nuclear RNA-Binding Proteins
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Krainer A R
Cold Spring Harbor Laboratory, NY 11724.
References (53)
53 references, click to expand
  1. A compensatory base change in U1 snRNA suppresses a 5' splice site mutation.
    Cell. 1986 Sep 12;46(6):827-35 PMID: 3757028
  2. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  3. A protein associated with small nuclear ribonucleoprotein particles recognizes the 3' splice site of premessenger RNA.
    Cell. 1986 Dec 26;47(6):973-84 PMID: 2946421
  4. RNA-protein organization of U1, U5 and U4-U6 small nuclear ribonucleoproteins in HeLa cells.
    J Mol Biol. 1986 Jun 5;189(3):519-32 PMID: 2946870
  5. Catalysis by RNA.
    Bioessays. 1986 Feb;4(2):56-61 PMID: 2431683
  6. Pre-mRNA splicing.
    Annu Rev Genet. 1986;20:671-708 PMID: 2880558
  7. Purification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents.
    EMBO J. 1986 Dec 20;5(13):3509-16 PMID: 2951249
  8. Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA.
    Cell. 1987 Apr 24;49(2):229-39 PMID: 3552247
  9. An essential yeast snRNA with a U5-like domain is required for splicing in vivo.
    Cell. 1987 Jun 5;49(5):613-24 PMID: 3555841
  10. Fractionation of HeLa cell nuclear extracts reveals minor small nuclear ribonucleoprotein particles.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8408-12 PMID: 2960976
  11. Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions.
    Cell. 1987 Dec 24;51(6):1019-26 PMID: 2961458
  12. Separation of multiple components of HeLa cell nuclear extracts required for pre-messenger RNA splicing.
    J Biol Chem. 1987 Dec 25;262(36):17630-40 PMID: 2961739
  13. The ribosome returns.
    Nature. 1988 Jan 21;331(6153):223-7 PMID: 3275901
  14. In vitro assembly of U1 snRNPs.
    EMBO J. 1987 Nov;6(11):3479-85 PMID: 2962858
  15. Reconstitution of the U1 small nuclear ribonucleoprotein particle.
    Mol Cell Biol. 1987 Nov;7(11):4030-7 PMID: 2963210
  16. Each of the activities of signal recognition particle (SRP) is contained within a distinct domain: analysis of biochemical mutants of SRP.
    Cell. 1988 Jan 15;52(1):39-49 PMID: 2830980
  17. Requirement of ATP in the second step of the pre-mRNA splicing reaction.
    Nucleic Acids Res. 1988 Apr 25;16(8):3157-64 PMID: 3375053
  18. Pre-mRNA splicing in vitro requires intact U4/U6 small nuclear ribonucleoprotein.
    Cell. 1986 Aug 29;46(5):697-704 PMID: 2427202
  19. Fractionation and characterization of human small nuclear ribonucleoproteins containing U1 and U2 RNAs.
    J Biol Chem. 1983 Jun 10;258(11):7181-9 PMID: 6189835
  20. Polypeptide components of human small nuclear ribonucleoproteins.
    Proc Natl Acad Sci U S A. 1983 Sep;80(17):5208-12 PMID: 6225117
  21. Splicing of messenger RNA precursors is inhibited by antisera to small nuclear ribonucleoprotein.
    Cell. 1983 Nov;35(1):101-7 PMID: 6194895
  22. 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
  23. Immunoelectron microscopic localization of snRNPs.
    Biol Cell. 1983;49(1):1-10 PMID: 6230127
  24. Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro.
    Cell. 1984 Apr;36(4):993-1005 PMID: 6323033
  25. U4 and U6 RNAs coexist in a single small nuclear ribonucleoprotein particle.
    Nucleic Acids Res. 1984 Apr 11;12(7):3283-93 PMID: 6201826
  26. 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
  27. Anti-RNP monoclonal antibodies derived from a mouse strain with lupus-like autoimmunity.
    J Immunol. 1982 Mar;128(3):1176-80 PMID: 6173424
  28. A rapid, novel method for the solid-phase derivatization of IgG antibodies for immune-affinity chromatography.
    J Immunol Methods. 1978;24(3-4):305-9 PMID: 82592
  29. Are snRNPs involved in splicing?
    Nature. 1980 Jan 10;283(5743):220-4 PMID: 7350545
  30. A mechanism for RNA splicing.
    Proc Natl Acad Sci U S A. 1980 Apr;77(4):1877-9 PMID: 6246511
  31. Silver staining of proteins in polyacrylamide gels.
    Anal Biochem. 1981 Nov 15;118(1):197-203 PMID: 6175245
  32. Antibodies to RNA from autoimmune NZB/NZW mice recognize a similar antigenic determinant and show a large idiotypic diversity.
    Proc Natl Acad Sci U S A. 1982 Jun;79(12):3818-22 PMID: 6179087
  33. A one-step purification of membrane proteins using a high efficiency immunomatrix.
    J Biol Chem. 1982 Sep 25;257(18):10766-9 PMID: 6955305
  34. Isolation of small nuclear ribonucleoproteins containing U1, U2, U4, U5, and U6 RNAs.
    J Biol Chem. 1983 Feb 25;258(4):2604-13 PMID: 6185498
  35. Isolation and characterization of rabbit anti-m3 2,2,7G antibodies.
    Nucleic Acids Res. 1982 Nov 25;10(22):7103-13 PMID: 7155893
  36. Evidence for the existence of snRNAs U4 and U6 in a single ribonucleoprotein complex and for their association by intermolecular base pairing.
    EMBO J. 1984 Jun;3(6):1357-63 PMID: 6204860
  37. The 5' terminus of the RNA moiety of U1 small nuclear ribonucleoprotein particles is required for the splicing of messenger RNA precursors.
    Cell. 1984 Aug;38(1):299-307 PMID: 6235919
  38. 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
  39. Characterization of U small nuclear RNA-associated proteins.
    J Biol Chem. 1984 Oct 25;259(20):12850-6 PMID: 6208190
  40. Primary and secondary structure of U8 small nuclear RNA.
    J Biol Chem. 1985 Sep 15;260(20):10930-5 PMID: 2411727
  41. 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
  42. U2 as well as U1 small nuclear ribonucleoproteins are involved in premessenger RNA splicing.
    Cell. 1985 Oct;42(3):737-50 PMID: 2996775
  43. The 3' splice site of pre-messenger RNA is recognized by a small nuclear ribonucleoprotein.
    Science. 1985 Dec 20;230(4732):1344-9 PMID: 2933810
  44. Purification of a protein required for the splicing of pre-mRNA and its separation from the lariat debranching enzyme.
    EMBO J. 1985 Dec 16;4(13A):3571-81 PMID: 4092689
  45. Specific conjugation reactions of the oligosaccharide moieties of immunoglobulins.
    J Appl Biochem. 1985 Aug-Oct;7(4-5):347-55 PMID: 3912370
  46. RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):887-91 PMID: 3456573
  47. Heterogeneous nuclear ribonucleoproteins: role in RNA splicing.
    Science. 1986 Mar 28;231(4745):1534-9 PMID: 3952495
  48. Separation of IgG idiotypes by high-performance hydroxylapatite chromatography.
    Methods Enzymol. 1986;121:615-22 PMID: 3014269
  49. Splicing of messenger RNA precursors.
    Annu Rev Biochem. 1986;55:1119-50 PMID: 2943217
  50. U1, U2, and U4/U6 small nuclear ribonucleoproteins are required for in vitro splicing but not polyadenylation.
    Cell. 1986 Aug 29;46(5):691-6 PMID: 2427201
  51. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  52. The derivatization of oxidized polysaccharides for protein immobilization and affinity chromatography.
    Biochim Biophys Acta. 1976 Mar 25;428(1):157-65 PMID: 1260016
  53. A protein that specifically recognizes the 3' splice site of mammalian pre-mRNA introns is associated with a small nuclear ribonucleoprotein.
    Cell. 1986 Dec 5;47(5):755-66 PMID: 2946417
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/16.20.9415
Published
1988-10-25
Pages
9415-29
Language
English
Region
England
NLM ID
0411011
PMCID
PMC338753
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/16.20.9415 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: [email protected]

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