Home LiteratureArticle Details
PMID: 8265352 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of U3 snoRNP from CHO cells: a novel 55 kDa protein binds to the central part of U3 snoRNA.

Nucleic acids research ·Vol. 21 ·No. 23 ·1993-11-25 ·Pages 5377-85

Lübben B, Marshallsay C, Rottmann N, Lührmann R

Abstract

U3 snoRNP, the most abundant of the small nucleolar ribonucleoprotein particles (snoRNPs), has previously been demonstrated to participate in pre-rRNA maturation. Here we report the purification of U3 snoRNP from CHO cells using anti-m3G-immunoaffinity and mono Q anion-exchange chromatography. Isolated U3 snoRNPs contain three novel proteins, of 15, 50 and 55 kDa respectively. These proteins may represent core U3 snoRNP proteins whose binding mediates the association of other proteins, such as fibrillarin, that are lost during purification. Using a rabbit antiserum raised against the 55 kDa protein, and an in vitro reconstitution assay, we have localised the 55 kDa protein binding site on the U3 snoRNA. Stable binding of the 55 kDa protein requires sequences located between nucleotides 97 and 204 of the human U3 snoRNA, including the evolutionarily conserved B and C sequence motifs.

MeSH Terms
Animals Base Sequence Binding Sites CHO Cells Cell Nucleolus/chemistry Cricetinae Molecular Sequence Data Molecular Weight Nuclear Proteins/chemistry Nucleic Acid Conformation RNA, Small Nuclear/metabolism Ribonucleoproteins, Small Nuclear/chemistry Structure-Activity Relationship
Chemicals
Nuclear Proteins RNA, Small Nuclear Ribonucleoproteins, Small Nuclear
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lübben B
Institut für Molekularbiologie und Tumorforschung, Phillipps-Universität Marburg, Germany.
Marshallsay C
Rottmann N
Lührmann R
References (29)
29 references, click to expand
  1. Structure and function of nucleolar snRNPs.
    Mol Biol Rep. 1993 Aug;18(2):149-56 PMID: 7694081
  2. The small nucleolar RNP protein NOP1 (fibrillarin) is required for pre-rRNA processing in yeast.
    EMBO J. 1991 Mar;10(3):573-83 PMID: 1825809
  3. 3'-terminal labelling of RNA with T4 RNA ligase.
    Nature. 1978 Oct 12;275(5680):560-1 PMID: 692735
  4. 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
  5. Maturation of a 100 kDa protein associated with preribosomes in Chinese hamster ovary cells.
    Mol Biol Rep. 1983 May;9(1-2):39-47 PMID: 6350846
  6. 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
  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. The yeast homologue of U3 snRNA.
    EMBO J. 1987 Jul;6(7):2145-55 PMID: 3308452
  9. A monoclonal antibody against 2,2,7-trimethylguanosine that reacts with intact, class U, small nuclear ribonucleoproteins as well as with 7-methylguanosine-capped RNAs.
    Eur J Biochem. 1987 Oct 15;168(2):461-7 PMID: 2959477
  10. Structural analysis of the human U3 ribonucleoprotein particle reveal a conserved sequence available for base pairing with pre-rRNA.
    Mol Cell Biol. 1987 Aug;7(8):2899-913 PMID: 2959855
  11. Structure and organization of mouse U3B RNA functional genes.
    J Biol Chem. 1988 Dec 25;263(36):19461-7 PMID: 3198636
  12. Genes for human U3 small nucleolar RNA contain highly conserved flanking sequences.
    Biochim Biophys Acta. 1989 Jun 1;1008(1):14-22 PMID: 2719960
  13. U3, U8 and U13 comprise a new class of mammalian snRNPs localized in the cell nucleolus.
    EMBO J. 1989 Oct;8(10):3113-9 PMID: 2531075
  14. A yeast nucleolar protein related to mammalian fibrillarin is associated with small nucleolar RNA and is essential for viability.
    EMBO J. 1989 Dec 20;8(13):4015-24 PMID: 2686980
  15. The U3 small nucleolar ribonucleoprotein functions in the first step of preribosomal RNA processing.
    Cell. 1990 Mar 23;60(6):897-908 PMID: 2156625
  16. In vivo disruption of Xenopus U3 snRNA affects ribosomal RNA processing.
    EMBO J. 1990 Jul;9(7):2299-308 PMID: 2357971
  17. Amplification of plant U3 and U6 snRNA gene sequences using primers specific for an upstream promoter element and conserved intragenic regions.
    Nucleic Acids Res. 1990 Jun 25;18(12):3459-66 PMID: 2362803
  18. Purification of small nuclear ribonucleoprotein particles with antibodies against modified nucleosides of small nuclear RNAs.
    Methods Enzymol. 1990;181:232-57 PMID: 2143251
  19. Structure of spliceosomal snRNPs and their role in pre-mRNA splicing.
    Biochim Biophys Acta. 1990 Nov 30;1087(3):265-92 PMID: 2147394
  20. Evidence for three distinct D proteins, which react differentially with anti-Sm autoantibodies, in the cores of the major snRNPs U1, U2, U4/U6 and U5.
    Nucleic Acids Res. 1990 Nov 25;18(22):6475-84 PMID: 1701240
  21. An intact Box C sequence in the U3 snRNA is required for binding of fibrillarin, the protein common to the major family of nucleolar snRNPs.
    EMBO J. 1991 Sep;10(9):2645-51 PMID: 1714385
  22. Depletion of U3 small nucleolar RNA inhibits cleavage in the 5' external transcribed spacer of yeast pre-ribosomal RNA and impairs formation of 18S ribosomal RNA.
    EMBO J. 1991 Dec;10(13):4231-9 PMID: 1756730
  23. Distinct molecular signals for nuclear import of the nucleolar snRNA, U3.
    Genes Dev. 1992 Jun;6(6):1120-30 PMID: 1592260
  24. Characterization of the U3 and U6 snRNA genes from wheat: U3 snRNA genes in monocot plants are transcribed by RNA polymerase III.
    Plant Mol Biol. 1992 Sep;19(6):973-83 PMID: 1511142
  25. In vitro reconstitution of U1 and U2 snRNPs from isolated proteins and snRNA.
    Mol Biol Rep. 1992 Sep;16(4):229-40 PMID: 1454055
  26. The nucleolar snRNAs: catching up with the spliceosomal snRNAs.
    Trends Biochem Sci. 1993 Apr;18(4):131-5 PMID: 8493724
  27. A U3 snoRNP protein with homology to splicing factor PRP4 and G beta domains is required for ribosomal RNA processing.
    EMBO J. 1993 Jun;12(6):2549-58 PMID: 8508778
  28. A small nucleolar RNA is processed from an intron of the human gene encoding ribosomal protein S3.
    Genes Dev. 1993 Jul;7(7A):1176-90 PMID: 8319909
  29. Isolation of nucleoli. A method that combines high yield, structural integrity, and biochemical preservation.
    J Cell Biol. 1971 Nov;51(21):563-8 PMID: 4939528
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.23.5377
Published
1993-11-25
Pages
5377-85
Language
English
Region
England
NLM ID
0411011
PMCID
PMC310574
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/21.23.5377 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