In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation.
10.1101/gad.3.4.479
Crossref journal-article
Cold Spring Harbor Laboratory
Genes & Development (246)
Abstract

We have reconstituted in vitro the four snRNPs known to be involved in pre-mRNA splicing: U1, U2, U5, and U4/6. Reconstitution involves adding either authentic or in vitro-synthesized snRNAs to extracts enriched in snRNP structural polypeptides. The reconstituted snRNPs have the same buoyant density and are immunoprecipitated by the same antibodies as authentic snRNPs. Thus, the polypeptide composition of reconstituted snRNPs is similar, if not identical, to that of authentic snRNPs. We show further that a reconstituted U4/U6 particle is fully functional in forming splicing complexes with pre-mRNA. As is the case for the authentic U4/U6 snRNP, the reconstituted U4 snRNP, but not the U6 snRNA, dissociates from the complex prior to formation of the mature spliceosome. The ability to reconstitute snRNPs and assay their activity in spliceosome formation should provide a powerful approach to study these particles.

Bibliography

Pikielny, C. W., Bindereif, A., & Green, M. R. (1989). In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation. Genes & Development, 3(4), 479–487.

Authors 3
  1. C W Pikielny (first)
  2. A Bindereif (additional)
  3. M R Green (additional)
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Dates
Type When
Created 18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited 2 years, 3 months ago (May 12, 2023, 2:22 a.m.)
Indexed 1 year, 10 months ago (Oct. 23, 2023, 11:02 a.m.)
Issued 36 years, 4 months ago (April 1, 1989)
Published 36 years, 4 months ago (April 1, 1989)
Published Online 36 years, 4 months ago (April 1, 1989)
Published Print 36 years, 4 months ago (April 1, 1989)
Funders 0

None

@article{Pikielny_1989, title={In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation.}, volume={3}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.3.4.479}, DOI={10.1101/gad.3.4.479}, number={4}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Pikielny, C W and Bindereif, A and Green, M R}, year={1989}, month=apr, pages={479–487} }