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Rab10 GTPase regulates ER dynamics and morphology

Nature Cell Biology volume 15, pages 169–178 (2013)Cite this article

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Abstract

We have identified Rab10 as an ER-specific Rab GTPase that regulates ER structure and dynamics. We show that Rab10 localizes to the ER and to dynamic ER-associated structures that track along microtubules and mark the position of new ER tubule growth. Rab10 depletion or expression of a Rab10 GDP-locked mutant alters ER morphology, resulting in fewer ER tubules. We demonstrate that this defect is due to a reduced ability of dynamic ER tubules to grow out and successfully fuse with adjacent ER. Consistent with this function, Rab10 partitions to dynamic ER-associated domains found at the leading edge of almost half of all dynamic ER tubules. Interestingly, this Rab10 domain is highly enriched with at least two ER enzymes that regulate phospholipid synthesis, phosphatidylinositol synthase (PIS) and CEPT1. Both the formation and function of this Rab10/PIS/CEPT1 dynamic domain are inhibited by expression of a GDP-locked Rab10 mutant. Together, these data demonstrate that Rab10 regulates ER dynamics and further suggest that these dynamics could be coupled to phospholipid synthesis.

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Figure 1: Purification of GTP-binding proteins from a X. laevis in vitro ER assembly assay.
Figure 2: Rab10 localizes to the ER and regulates tubular ER morphology.
Figure 3: Depletion of endogenous Rab10 reduces tubular ER morphology.
Figure 4: Rab10 regulates ER tubule dynamics and fusion.
Figure 5: Rab10 and PIS co-localize to the leading edge of dynamic ER tubules.
Figure 6: CEPT1 and PIS partition with Rab10 to ER-associated dynamic domains.
Figure 7: Rab10 regulates the formation of Rab10/PIS/CEPT1 dynamic puncta.
Figure 8: Atlastin and Rab10 do not overlap in function or location.

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Acknowledgements

We thank C. English for helpful suggestions, A. Merz (University of Washington, USA) for generously providing Rab GDI, and E. Snapp, T. Lee and J. Friedman for constructs. This work was supported by NIH grants GM083977 to G.K.V. and GM07135 to A.R.E.

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Authors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80309, USA

    Amber R. English & Gia K. Voeltz

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  1. Amber R. English
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Contributions

G.K.V. and A.R.E. designed the experiments and wrote the manuscript. A.R.E. performed the experiments and data analysis.

Corresponding author

Correspondence to Gia K. Voeltz.

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The authors declare no competing financial interests.

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Relative localization over time of Rab10 WT and an ER luminal protein.

Cos-7 cells expressing mCh-Rab10 WT and KDEL-venus (Rab10 WT, red; KDEL, green) were imaged live by confocal fluorescence microscopy. Scale bar, 10 μm. (AVI 9988 kb)

Relative localization of PIS, Rab10 WT and an ER luminal protein.

Cos-7 cells co-expressing mCh-PIS, BFP-Rab10 WT and KDEL-venus (PIS, red; Rab10 WT, blue; KDEL, green) were imaged live by confocal fluorescence microscopy. Scale bar, 10 μm. (AVI 23812 kb)

Relative localization of Rab10 WT, CEPT1 and PIS

Cos-7 cells co-expressing BFP-Rab10 WT, mCh-CEPT1 and GFP-PIS (Rab10 WT, blue; CEPT1, red, GFP, green) were imaged live by confocal fluorescence microscopy. Scale bar, 10 μm. (AVI 9988 kb)

Relative localization and structure of the ER in cells expressing Rab10 T23N, CEPT1 and PIS

Cos-7 cells co-expressing BFP-Rab10 T23N, mCh-CEPT1 and GFP-PIS (Rab10 T23N, blue; CEPT1, red, GFP, green) were imaged live by confocal fluorescence microscopy. Scale bar, 10 μm. (AVI 9988 kb)

Relative localization of PIS, Rab10 WT and Atl3 puncta

Cos-7 cells expressing mCh-PIS, BFP-Rab10 WT and GFP-Atl3 (mCh-PIS, red; BFPRab10 WT, blue; GFP-Atl3, green) were imaged live by confocal fluorescence microscopy. Scale bar, 10 μm. (AVI 14596 kb)

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English, A., Voeltz, G. Rab10 GTPase regulates ER dynamics and morphology. Nat Cell Biol 15, 169–178 (2013). https://doi.org/10.1038/ncb2647

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