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Inositol hexakisphosphate and Gle1 activate the DEAD-box protein Dbp5 for nuclear mRNA export

Nature Cell Biology volume 8, pages 711–716 (2006)Cite this article

Abstract

Regulation of nuclear mRNA export is critical for proper eukaryotic gene expression. A key step in this process is the directional translocation of mRNA–ribonucleoprotein particles (mRNPs) through nuclear pore complexes (NPCs) that are embedded in the nuclear envelope1. Our previous studies in Saccharomyces cerevisiae defined an in vivo role for inositol hexakisphosphate (InsP6) and NPC-associated Gle1 in mRNA export2. Here, we show that Gle1 and InsP6 act together to stimulate the RNA-dependent ATPase activity of the essential DEAD-box protein Dbp5. Overexpression of DBP5 specifically suppressed mRNA export and growth defects of an ipk1 nup42 mutant defective in InsP6 production and Gle1 localization. In vitro kinetic analysis showed that InsP6 significantly increased Dbp5 ATPase activity in a Gle1-dependent manner and lowered the effective RNA concentration for half-maximal ATPase activity. Gle1 alone had minimal effects. Maximal InsP6 binding required both Dbp5 and Gle1. It has been suggested that Dbp5 requires unidentified cofactors3,4. We now propose that Dbp5 activation at NPCs requires Gle1 and InsP6. This would facilitate spatial control of the remodelling of mRNP protein composition during directional transport and provide energy to power transport cycles.

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Figure 1: DBP5 overexpression rescues the mRNA export and growth defects of the ipk1Δ nup42Δ mutant.
Figure 2: InsP6 and Gle1 stimulate Dbp5 ATPase activity.
Figure 3: InsP6 binding in the presence of Gle1 and Dbp5.
Figure 4: Schematic representation of how Gle1–InsP6 activation of Dbp5 may trigger mRNP remodelling at a NPC cytoplasmic-face scaffold.

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Acknowledgements

We thank C. Cole, J. York, R. Emeson and M. Fournier for plasmids, strains and reagents, and J. York and members of the Wente and Hardy labs for critical discussions and comments. This work was supported by a National Institutes of Health (NIH) grant (R01–GM51219) to S.R.W. and an National Research Service Award (NSRA) NIH Virus, Nucleic Acids and Cancer position (5T32–CA009385) and Ruth Kirschstein NIH NSRA (1F32–GM075459) to E.J.T.

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  1. Department of Cell and Developmental Biology, Vanderbilt University Medical Center, U-3209 MRBIII, 465 21st Avenue South, Nashville, 37232–8240, TN, USA

    Abel R. Alcázar-Román, Elizabeth J. Tran, Shuangli Guo & Susan R. Wente

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Correspondence to Susan R. Wente.

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Alcázar-Román, A., Tran, E., Guo, S. et al. Inositol hexakisphosphate and Gle1 activate the DEAD-box protein Dbp5 for nuclear mRNA export. Nat Cell Biol 8, 711–716 (2006). https://doi.org/10.1038/ncb1427

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