A synthetic A tail rescues yeast nuclear accumulation of a ribozyme-terminated transcript

To investigate the role of 3' end formation in yeast mRNA export, we replaced the mRNA cleavage and polyadenylation signal with a self-cleaving hammerhead ribozyme element. The resulting RNA is unadenylated and accumulates near its site of synthesis. Nonetheless, a significant fraction of this...

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Bibliographic Details
Published in:RNA (Cambridge) 2004-12, Vol.10 (12), p.1888-1899
Main Authors: Dower, Ken, Kuperwasser, Nicolas, Merrikh, Houra, Rosbash, Michael
Format: Article
Language:English
Subjects:
Base Sequence
Cell Nucleus - metabolism
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
In Situ Hybridization, Fluorescence
Models, Biological
Mutation
Poly A - chemistry
Poly A - genetics
Poly A - metabolism
Poly(A)-Binding Proteins - genetics
Poly(A)-Binding Proteins - metabolism
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
RNA, Catalytic - metabolism
RNA, Fungal - chemistry
RNA, Fungal - genetics
RNA, Fungal - metabolism
RNA, Messenger - chemistry
RNA, Messenger - genetics
RNA, Messenger - metabolism
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
Transcription, Genetic
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Summary:To investigate the role of 3' end formation in yeast mRNA export, we replaced the mRNA cleavage and polyadenylation signal with a self-cleaving hammerhead ribozyme element. The resulting RNA is unadenylated and accumulates near its site of synthesis. Nonetheless, a significant fraction of this RNA reaches the cytoplasm. Nuclear accumulation was relieved by insertion of a stretch of DNA-encoded adenosine residues immediately upstream of the ribozyme element (a synthetic A tail). This indicates that a 3' stretch of adenosines can promote export, independently of cleavage and polyadenylation. We further show that a synthetic A tail-containing RNA is unaffected in 3' end formation mutant strains, in which a normally cleaved and polyadenylated RNA accumulates within nuclei. Our results support a model in which a polyA tail contributes to efficient mRNA progression away from the gene, most likely through the action of the yeast polyA-tail binding protein Pab1p.
ISSN:1355-8382
1469-9001
DOI:10.1261/rna.7166704