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PMID:21329879

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Citation

Nagaike, T, Logan, C, Hotta, I, Rozenblatt-Rosen, O, Meyerson, M and Manley, JL (2011) Transcriptional activators enhance polyadenylation of mRNA precursors. Mol. Cell 41:409-18

Abstract

Polyadenylation of mRNA precursors is frequently coupled to transcription by RNA polymerase II. Although this coupling is known to involve interactions with the C-terminal domain of the RNA polymerase II largest subunit, the possible role of other factors is not known. Here we show that a prototypical transcriptional activator, GAL4-VP16, stimulates transcription-coupled polyadenylation in vitro. In the absence of GAL4-VP16, specifically initiated transcripts accumulated but little polyadenylation was observed, while in its presence polyadenylation was strongly enhanced. We further show that this stimulation requires the transcription elongation-associated PAF complex (PAF1c), as PAF1c depletion blocked GAL4-VP16-stimulated polyadenylation. Furthermore, knockdown of PAF subunits by siRNA resulted in decreased 3' cleavage, and nuclear export, of mRNA in vivo. Finally, we show that GAL4-VP16 interacts directly with PAF1c and recruits it to DNA templates. Our results indicate that a transcription activator can stimulate transcription-coupled 3' processing and does so via interaction with PAF1c.

Links

PubMed PMC3060669 Online version:10.1016/j.molcel.2011.01.022

Keywords

Cells, Cultured; Humans; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Polyadenylation; RNA Polymerase II/genetics; RNA Polymerase II/metabolism; RNA Precursors/metabolism; RNA, Messenger/metabolism; RNA, Small Interfering/metabolism; Trans-Activators/genetics; Trans-Activators/metabolism; Transcription, Genetic; Transfection

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