Warning: Undefined array key "mm" in /www/wwwroot/www.ai-bt.com/si.php on line 10 Deprecated: trim(): Passing null to parameter #1 ($string) of type string is deprecated in /www/wwwroot/www.ai-bt.com/si.php on line 10 Antisense probing of the human U4/U6 snRNP with biotinylated 2'-OMe RNA oligonucleotides.

Literature DB >> 2478298

Antisense probing of the human U4/U6 snRNP with biotinylated 2'-OMe RNA oligonucleotides.

B J Blencowe¹, B S Sproat, U Ryder, S Barabino, A I Lamond.

Abstract

We have used antisense 2'-OMe RNA oligonucleotides carrying four 5'-terminal biotin residues to probe the structure and function of the human U4/U6 snRNP. Nine oligonucleotides, complementary to multiple regions of U4 and U6 snRNAs, bound stably and specifically to U4/U6 snRNP. This allowed for efficient and selective removal of U4/U6 from HeLa cell nuclear extracts. Binding of oligonucleotides to certain snRNA domains inhibited splicing and affected the U4-U6 interaction. Pre-mRNA and splicing products could also be affinity-selected through binding of the oligonucleotides to U4/U6 snRNPs in splicing complexes. The results suggest that U4 snRNP is not released during spliceosome assembly.

Entities: Chemical Gene Species

Mesh：

Substances：

Year: 1989 PMID： 2478298 DOI： 10.1016/0092-8674(89)90036-6

Source DB: PubMed Journal: Cell ISSN： 0092-8674 Impact factor: 41.582

Keyword Cloud
Cited

63 in total

1. The spliceosome assembly pathway in mammalian extracts.

Authors: S F Jamison; A Crow; M A Garcia-Blanco
Journal: Mol Cell Biol Date: 1992-10 Impact factor: 4.272

2. Purification of splicing factor SF1, a heat-stable protein that functions in the assembly of a presplicing complex.

Authors: A Krämer
Journal: Mol Cell Biol Date: 1992-10 Impact factor: 4.272

3. Multiple functional domains of human U2 small nuclear RNA: strengthening conserved stem I can block splicing.

Authors: J Wu; J L Manley
Journal: Mol Cell Biol Date: 1992-12 Impact factor: 4.272

4. Reconstitution of functional mammalian U4 small nuclear ribonucleoprotein: Sm protein binding is not essential for splicing in vitro.

Authors: C Wersig; A Bindereif
Journal: Mol Cell Biol Date: 1992-04 Impact factor: 4.272

5. Activity of chimeric RNAs of U6 snRNA and (-)sTRSV in the cleavage of a substrate RNA.

Authors: T Tani; Y Takahashi; Y Ohshima
Journal: Nucleic Acids Res Date: 1992-06-25 Impact factor: 16.971

6. Protein composition and electron microscopy structure of affinity-purified human spliceosomal B complexes isolated under physiological conditions.

Authors: Jochen Deckert; Klaus Hartmuth; Daniel Boehringer; Nastaran Behzadnia; Cindy L Will; Berthold Kastner; Holger Stark; Henning Urlaub; Reinhard Lührmann
Journal: Mol Cell Biol Date: 2006-07 Impact factor: 4.272

Antisense probing of the human U4/U6 snRNP with biotinylated 2'-OMe RNA oligonucleotides.

1. The spliceosome assembly pathway in mammalian extracts.

2. Purification of splicing factor SF1, a heat-stable protein that functions in the assembly of a presplicing complex.

3. Multiple functional domains of human U2 small nuclear RNA: strengthening conserved stem I can block splicing.

4. Reconstitution of functional mammalian U4 small nuclear ribonucleoprotein: Sm protein binding is not essential for splicing in vitro.

5. Activity of chimeric RNAs of U6 snRNA and (-)sTRSV in the cleavage of a substrate RNA.

6. Protein composition and electron microscopy structure of affinity-purified human spliceosomal B complexes isolated under physiological conditions.

7. Functional spliceosomal A complexes can be assembled in vitro in the absence of a penta-snRNP.

8. Antisense probes containing 2-aminoadenosine allow efficient depletion of U5 snRNP from HeLa splicing extracts.

9. Purification of RNA and RNA-protein complexes by an R17 coat protein affinity method.

Review 10. Insight into lncRNA biology using hybridization capture analyses.