Dynamics of single mRNP nucleocytoplasmic transport and export through the nuclear pore in living cells

Amir Mor, Shimrit Suliman, Rakefet Ben-Yishay, Sharon Yunger, Yehuda Brody, Yaron Shav-Tal

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204 Scopus citations

Abstract

The flow of genetic information in eukaryotic cells occurs through the nucleocytoplasmic translocation of mRNAs. Knowledge of in vivo messenger RNA export kinetics remains poor in comparison with that of protein transport. We have established a mammalian system that allowed the real-time visualization and quantification of large single mRNA-protein complexes (mRNPs) during export. The in vivo dynamics of bulk mRNP transport and export, from transcription to the nuclear pore complex (NPC), occurred within a 5-40 minute time frame, with no NPC pile-up. mRNP export was rapid (about 0.5s) and kinetically faster than nucleoplasmic diffusion. Export inhibition demonstrated that mRNA-NPC interactions were independent of ongoing export. Nucleoplasmic transport dynamics of intron-containing and intronless mRNAs were similar, yet an intron did increase export efficiency. Here we provide visualization and analysis at the single mRNP level of the various steps in nuclear gene expression and the inter-chromatin tracks through which mRNPs diffuse, and demonstrate the kinetics of mRNP-NPC interactions and translocation.

Original languageEnglish
Pages (from-to)543-552
Number of pages10
JournalNature Cell Biology
Volume12
Issue number6
DOIs
StatePublished - Jun 2010

Bibliographical note

Funding Information:
We thank J. Tremblay (Laval University, Canada) for GFP–Dys and GFP–mini-Dys, J. Ellenberg (EMBL Heidelberg, Germany) for the POM121-mCherry plasmid, and R. Drummer (Bar-Ilan University (BIU), Israel) for statistical analysis. This work was supported by grants to Y.S.T. from the Israel Science Foundation (grant 250/06) and the Israel Ministries of Health and Science. Y.S.T. thanks the Israel Science Foundation for the fluorescence live-cell imaging microscope. Y.B. is an Azrielli fellow. Y.S.T. is the Jane Stern Lebell Family Fellow in Life Sciences at BIU.

Funding

We thank J. Tremblay (Laval University, Canada) for GFP–Dys and GFP–mini-Dys, J. Ellenberg (EMBL Heidelberg, Germany) for the POM121-mCherry plasmid, and R. Drummer (Bar-Ilan University (BIU), Israel) for statistical analysis. This work was supported by grants to Y.S.T. from the Israel Science Foundation (grant 250/06) and the Israel Ministries of Health and Science. Y.S.T. thanks the Israel Science Foundation for the fluorescence live-cell imaging microscope. Y.B. is an Azrielli fellow. Y.S.T. is the Jane Stern Lebell Family Fellow in Life Sciences at BIU.

FundersFunder number
Israel Ministries of Health and Science
Israel Science Foundation250/06

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