Identification of Prp40, a novel essential yeast splicing factor associated with the U1 small nuclear ribonucleoprotein particle

Hung Ying Kao, Paul G Siliciano

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75 Scopus citations

Abstract

We have used suppressor genetics to identify factors that interact with Saccharomyces cerevisiae U1 small nuclear RNA (snRNA). In this way, we isolated PRP40-1, a suppressor that restores growth at 18°C to a strain bearing a cold-sensitive mutation in U1 RNA. A gene disruption experiment shows that PRP40 is an essential gene. To study the role of PRP40 in splicing, we created a pool of temperature-sensitive prp40 strains. Primer extension analysis of intron-containing transcripts in prp40 temperature- sensitive strains reveals a splicing defect, indicating that Prp40 is a general splicing factor. In vitro heat inactivation and complementation assays demonstrate that Prp40 plays a direct role in pre-mRNA splicing. In addition, U1 RNA coimmunoprecipitates with Prp40, indicating that Prp40 is bound to the U1 small nuclear ribonucleoprotein particle in vivo. Therefore, we conclude that PRP40 encodes a novel, essential splicing component that associates with the yeast U1 small nuclear ribonucleoprotein particle.

Original languageEnglish (US)
Pages (from-to)960-967
Number of pages8
JournalMolecular and cellular biology
Volume16
Issue number3
DOIs
StatePublished - Mar 1996

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