A Nuclear Export Block Triggers the Decay of Newly Synthesized Polyadenylated RNA
Agnieszka Tudek1, Manfred Schmid1, Marius Makaras1
1Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, Building 1130, 8000 Aarhus C, Denmark.
Abstract:
Genomes are promiscuously transcribed, necessitating mechanisms that facilitate the sorting of RNA for function or destruction. The polyA (pA) tail is one such distinguishing feature, which in the Saccharomyces cerevisiae nucleus is bound by the Nab2p protein, yielding transcript protection. As Nab2p also contacts the main nuclear export factor Mex67p, we asked whether transport kinetics contributes to RNA sorting. Indeed, 3' end sequencing of newly transcribed pA+ RNAs demonstrates that nuclear depletion of Mex67p elicits their instant and global decay. A similar phenotype is evident upon inactivation of other export factors and proportional to the amount of nuclear pA+ RNA. As RNA expression is partially rescued by Nab2p overexpression, we propose that an export block out-titrates Nab2p onto nuclear-retained pA+ RNA, reducing the pool of Nab2p available to protect new transcripts. More generally, we suggest that nuclear RNA decay, negotiated by Nab2p availability, aids in balancing cellular transcript supply with demand.
Insights
Nuclear RNA export is crucial for transcript stability. Blocking export factors like Mex67p causes rapid decay of polyadenylated (pA+) RNAs, suggesting a link between RNA transport and cellular RNA homeostasis.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Cells must regulate RNA levels due to extensive transcription.
- The polyadenylated (pA) tail is a key RNA feature.
- Nab2p protein binds pA tails in the nucleus, protecting RNA.
Purpose of the Study:
- To investigate if RNA transport kinetics influences RNA sorting.
- To determine the role of nuclear export factor Mex67p in RNA stability.
Main Methods:
- 3' end sequencing of newly transcribed polyadenylated (pA+) RNAs.
- Nuclear depletion of Mex67p and other export factors.
- Nab2p overexpression experiments.
Main Results:
- Depletion of Mex67p caused immediate and widespread decay of nuclear pA+ RNAs.
- Inactivation of other export factors showed similar decay phenotypes.
- Nab2p overexpression partially rescued RNA expression, suggesting competition for Nab2p.
Conclusions:
- Nuclear RNA export is critical for pA+ RNA stability.
- An export block can lead to RNA decay by out-titrating Nab2p.
- Nuclear RNA decay, regulated by Nab2p availability, balances cellular RNA supply and demand.
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