Single-molecule imaging of transcriptionally coupled and uncoupled splicing

Document Type

Article

Publication Date

11-23-2011

Abstract

Introns are removed from pre-mRNAs during transcription while the pre-mRNA is still tethered to the gene locus via RNA polymerase. However, during alternative splicing, it is important that splicing be deferred until all of the exons and introns involved in the choice have been synthesized. We have developed an in situ RNA imaging method with single-molecule sensitivity to define the intracellular sites of splicing. Using this approach, we found that the normally tight coupling between transcription and splicing is broken in situations where the intron's polypyrimidine tract is sequestered within strong secondary structures. We also found that in two cases of alternative splicing, in which certain exons are skipped due to the activity of the RNA-binding proteins Sxl and PTB, splicing is uncoupled from transcription. This uncoupling occurs only on the perturbed introns, whereas the preceding and succeeding introns are removed cotranscriptionally. © 2011 Elsevier Inc.

Identifier

81855183655 (Scopus)

Publication Title

Cell

External Full Text Location

https://doi.org/10.1016/j.cell.2011.10.024

e-ISSN

10974172

ISSN

00928674

PubMed ID

22118462

First Page

1054

Last Page

1065

Issue

5

Volume

147

Grant

MH 079197

Fund Ref

National Institutes of Health

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