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

doi:10.1038/ncb2056

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

Bar-Ilan University

Research output: Contribution to journal › Article › peer-review

223 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 language	English
Pages (from-to)	543-552
Number of pages	10
Journal	Nature Cell Biology
Volume	12
Issue number	6
DOIs	https://doi.org/10.1038/ncb2056
State	Published - 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.

Funders	Funder number
Israel Ministries of Health and Science
Israel Science Foundation	250/06

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10.1038/ncb2056

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title = "Dynamics of single mRNP nucleocytoplasmic transport and export through the nuclear pore in living cells",

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.",

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

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.",

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

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