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Activation of the DExD/H-box protein Dbp5 by the nuclear-pore protein Gle1 and its coactivator InsP6 is required for mRNA export

Nature Cell Biology volume 8, pages 668–676 (2006)Cite this article

Abstract

The DExD/H-box ATPase Dbp5 is essential for nuclear mRNA export, although its precise role in this process remains poorly understood. Here, we identify the nuclear pore protein Gle1 as a cellular activator of Dbp5. Dbp5 alone is unable to stably bind RNA or effectively hydrolyse ATP under physiological conditions, but addition of Gle1 dramatically stimulates these activities. A gle1 point mutant deficient for Dbp5 stimulation in vitro displays an mRNA export defect in vivo, indicating that activation of Dbp5 is an essential function of Gle1. Interestingly, Gle1 binds directly to inositol hexakisphosphate (InsP6) and InsP6 potentiates the Gle1-mediated stimulation of Dbp5. Dominant mutations in DBP5 and GLE1 that rescue mRNA export phenotypes associated with the lack of InsP6 mimic the InsP6 effects in vitro. Our results define specific functions for Gle1 and InsP6 in mRNA export and suggest that local activation of Dbp5 at the nuclear pore is critical for mRNA export.

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Figure 1: Gle1 is an activator of Dbp5 in vitro.
Figure 2: A novel temperature-sensitive gle1 allele identified through directed mutagenesis.
Figure 3: gle1R287A causes an mRNA export in vivo.
Figure 4: InsP6 modulates the interaction between Gle1 and Dbp5.
Figure 5: A dominant allele of DBP5 bypasses the requirement for InsP6 in vivo.
Figure 6: Dbp5L327V and Gle1H337R partially mimic the InsP6-mediated enhancement of kcat in vitro.

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Acknowledgements

We are grateful to P. Preker, C. Guthrie, F. Stutz, Z. Liu, and A.M. Pyle for providing reagents. We thank N. Pokala for the plasmids pSV271, pSV272 and pSV212. We also thank E. Jankowsky, A. Schoeffler, P. Kalab and M. Blower and other members of the Berger, Thorner and Weis labs for discussions and comments on the manuscript. This work was supported by an National Science Foundation predoctoral fellowship to C.S.W., by a National Scientist Development Grant from the American Heart Association to J.S.F. and by National Institutes of Health Research Grants (GM71747 to J.M.B., GM21841 to J.T. and GM58065 to K.W.).

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Author notes

  1. Jeffrey S. Flick

    Present address: Myriad Pharmaceuticals Inc., 320 Wakara Way, Salt Lake City, UT 84108, USA

Authors and Affiliations

  1. Division of Cell and Developmental Biology, University of California, Berkeley, Berkeley, 94720-3200, CA, USA

    Christine S. Weirich & Karsten Weis

  2. Division of Biochemistry and Molecular Biology, Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, 94720-3200, CA, USA

    Jan P. Erzberger, James M. Berger & Jeremy Thorner

  3. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, 37232-0146, TN, USA

    Jeffrey S. Flick

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Correspondence to Karsten Weis.

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Supplementary Figures S1, S2, S3, S4, S5, S6, Supplementary Tables S1, S2, S3 and Supplementary Methods (PDF 802 kb)

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Weirich, C., Erzberger, J., Flick, J. et al. Activation of the DExD/H-box protein Dbp5 by the nuclear-pore protein Gle1 and its coactivator InsP6 is required for mRNA export. Nat Cell Biol 8, 668–676 (2006). https://doi.org/10.1038/ncb1424

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