Abstract
We have investigated whether transcriptional activators influence the efficiency of constitutive splicing and 3'-end formation, in addition to transcription levels. Remarkably, strong activators result in higher levels of splicing and 3'-cleavage than weak activators and can control the efficiency of these steps in pre-mRNA processing separately. The pre-mRNA processing stimulatory property of activators is dependent on their binding to promoters, but is not an indirect consequence of the levels of transcripts produced. Moreover, stimulation of splicing and cleavage by a strong activator operates by a mechanism that requires the carboxyl-terminal domain of RNA polymerase II. The splicing stimulatory property of activators was observed for unrelated transcripts and for separate introns within a transcript, indicating a possible general role for strong activators in facilitating pre-mRNA processing levels. The results suggest that the efficiency of constitutive splicing and 3'-end cleavage is closely coordinated with transcription levels by promoter-bound activators.
Highlights
The generation of mature mRNA involves the addition of an m7G cap at the 5Ј-end, splicing, and cleavage and polyadenylation at the 3Ј-end of precursormRNAs
We have investigated whether transcriptional activators influence the efficiency of constitutive splicing and 3-end formation, in addition to transcription levels
These steps can occur in isolation of each other, the majority of pre-mRNA processing occurs as RNA polymerase II1 is transcribing
Summary
The generation of mature mRNA involves the addition of an m7G cap at the 5Ј-end, splicing, and cleavage and polyadenylation at the 3Ј-end of precursor (pre-)mRNAs. Our results demonstrate that the increased level of splicing and cleavage promoted by a strong activator is dependent on the CTD of pol II, indicating an important role for this domain in mediating the effects of promoters on pre-mRNA processing.
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