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Intracellular lumen extension requires ERM-1-dependent apical membrane expansion and AQP-8-mediated flux

An Erratum to this article was published on 01 March 2013

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Abstract

Many unicellular tubes such as capillaries form lumens intracellularly, a process that is not well understood. Here we show that the cortical membrane organizer ERM-1 is required to expand the intracellular apical/lumenal membrane and its actin undercoat during single-cell Caenorhabditis elegans excretory canal morphogenesis. We characterize AQP-8, identified in an ERM-1-overexpression (ERM-1[++]) suppressor screen, as a canalicular aquaporin that interacts with ERM-1 in lumen extension in a mercury-sensitive manner, implicating water-channel activity. AQP-8 is transiently recruited to the lumen by ERM-1, co-localizing in peri-lumenal cuffs interspaced along expanding canals. An ERM-1[++]-mediated increase in the number of lumen-associated canaliculi is reversed by AQP-8 depletion. We propose that the ERM-1/AQP-8 interaction propels lumen extension by translumenal flux, suggesting a direct morphogenetic effect of water-channel-regulated fluid pressure.

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Figure 1: ERM-1 is required to expand the excretory-canal lumenal membrane and its actin undercoat.
Figure 2: The ERM-1[++] cystic-canal phenotype is suppressed by loss of AQP-8.
Figure 3: AQP-8 promotes excretory canal lumen expansion and localizes to canalicular vesicles.
Figure 4: erm-1 and aqp-8 genetically interact in intracellular lumen morphogenesis.
Figure 5: AQP-8 and ERM-1 transiently co-localize during excretory canal development and physically interact in yeast two-hybrid assays.
Figure 6: AQP-8 functions as a water channel in canal morphogenesis.
Figure 7: ERM-1 recruits AQP-8 to the lumen and increases the canalicular–lumenal membrane connection.
Figure 8: Tomographic analysis of the ERM-1[++] effect on the canalicular–lumenal interface and a model of the ERM-1/AQP-8 function in excretory canal lumen extension.

Change history

  • 11 February 2013

    In the version of this Article that was originally published, the results section should have read: "Suppression of aqp-8(RNAi) was confirmed in..." The caption for Fig. 1il should have read: "ERM-1 dose-dependently restricts canal extension." The caption for Fig. 1o should have read: "1/4-extended canal with aligning vacuoles."

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Acknowledgements

We thank D. Baillie (Simon Fraser University, Burnaby, British Columbia, Canada), M. Futai (Osaka University, Osaka, Japan), M. Labouesse (IGBMC, France), K. Nehrke (University of Rochester Medical Center, Rochester New York, USA) and J. Simske (Case Western Reserve University School of Medicine, Cleveland, Ohio, USA), and the following C. elegans resource centres: J. Kohara (National Institute of Genetics, Mishima, Japan), S. Mitani (National Bioresource Project, Tokyo Women’s Medical University, Tokyo, Japan), the International C. elegans Gene Knockout Consortium and the Caenorhabditis elegans genetic centre (NIH Center for Research Resources) for providing plasmids and strains. We thank E. Membreno and D. Fernandez for technical support, A. Sengupta for three-dimensional graphics, A. Kim for image editing, F. Solomon for critical reading of the manuscript, and H. Weinstein and A. Walker for ongoing support. This work was supported by NIH grant GM078653 and a Mattina R. Proctor Award to V.G.

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L.A.K. performed experiments, generated, analysed and assembled most of the data and contributed to project design and manuscript writing. H.Z. and N.A. contributed to the generation of transgenic strains and RNAi experiments. L.S. and D.H.H. performed the TEM and tomographic analyses, and J.T.F. contributed to TEM experiments. M.B. generated the canal-specific endosomal marker strains, and J.T.F. and M.B. contributed to writing of the manuscript. V.G. conceived and directed the project, participated in experiments and wrote the manuscript.

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Correspondence to Verena Gobel.

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Khan, L., Zhang, H., Abraham, N. et al. Intracellular lumen extension requires ERM-1-dependent apical membrane expansion and AQP-8-mediated flux. Nat Cell Biol 15, 143–156 (2013). https://doi.org/10.1038/ncb2656

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