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RESEARCH PAPER
Departments of Biochemistry and Genetics and the Norris Cotton Cancer Center, Dartmouth Medical School, Hanover, New Hampshire 03755, USA
In eukaryotic cells, pre-mRNAs undergo extensive processing in the nucleus prior to export. Processing is subject to a quality-control mechanism that retains improperly processed transcripts at or near sites of transcription. A poly(A) tail added by the normal 3'-processing machinery is necessary but not sufficient for export. Retention depends on the exosome. In this study, we identify the poly(A)-binding protein, Pab1, and the poly(A) nuclease, PAN, as important factors that couple 3' processing to export. Pab1 contains a nonessential leucine-rich nuclear export signal and shuttles between the nucleus and the cytoplasm. It can exit the nucleus either as cargo of exportin 1 or bound to mRNA. Pab1 is essential but several bypass suppressors have been identified. Deletion of PAB1 from these bypass suppressor strains results in exosome-dependent retention at sites of transcription. Retention is also seen in cells lacking PAN, which Pab1 is thought to recruit and which may be responsible for the final step of mRNA biogenesis, trimming of the poly(A) tail to the length found on newly exported mRNAs. The studies presented here suggest that proper loading of Pab1 onto mRNAs and final trimming of the tail allows release from transcription sites and couples pre-mRNA processing to export.
[Keywords: mRNA transport; polyadenylation; exportin 1; nuclear export signal; gene expression; mRNA biogenesis]
Received September 28, 2004; revised version accepted November 2, 2004.
Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/gad.1267005.
1 These authors contributed equally to this work.
E-MAIL: charles.cole{at}dartmouth.edu; FAX (603) 650-1128.
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