Journalartikel

In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation.


AutorenlistePikielny, CW; Bindereif, A; Green, MR

Jahr der Veröffentlichung1989

Seiten479-487

ZeitschriftGenes & Development

Bandnummer3

Heftnummer4

ISSN0890-9369

eISSN1549-5477

DOI Linkhttps://doi.org/10.1101/gad.3.4.479

VerlagCold Spring Harbor Laboratory Press


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.



Zitierstile

Harvard-ZitierstilPikielny, C., Bindereif, A. and Green, M. (1989) In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation., Genes & Development, 3(4), pp. 479-487. https://doi.org/10.1101/gad.3.4.479

APA-ZitierstilPikielny, C., Bindereif, A., & Green, M. (1989). In vitro reconstitution of snRNPs: a reconstituted U4/U6 snRNP participates in splicing complex formation.. Genes & Development. 3(4), 479-487. https://doi.org/10.1101/gad.3.4.479


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