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Spatial control of actin organization at adherens junctions by a synaptotagmin-like protein

Nature volume 442, pages 580–584 (2006)Cite this article

Abstract

Epithelial tissues maintain a robust architecture during development. This fundamental property relies on intercellular adhesion through the formation of adherens junctions containing E-cadherin molecules1,2. Localization of E-cadherin is stabilized through a pathway involving the recruitment of actin filaments by E-cadherin3,4,5,6. Here we identify an additional pathway that organizes actin filaments in the apical junctional region (AJR) where adherens junctions form in embryonic epithelia. This pathway is controlled by Bitesize (Btsz), a synaptotagmin-like protein7,8 that is recruited in the AJR independently of E-cadherin and is required for epithelial stability in Drosophila embryos. On loss of btsz, E-cadherin is recruited normally to the AJR, but is not stabilized properly and actin filaments fail to form a stable continuous network. In the absence of E-cadherin, actin filaments are stable for a longer time than they are in btsz mutants. We identify two polarized cues that localize Btsz: phosphatidylinositol (4,5)-bisphosphate, to which Btsz binds; and Par-3. We show that Btsz binds to the Ezrin–Radixin–Moesin protein Moesin, an F-actin-binding protein that is localized apically9 and is recruited in the AJR in a btsz-dependent manner. Expression of a dominant-negative form of Ezrin that does not bind F-actin phenocopies the loss of btsz. Thus, our data indicate that, through their interaction, Btsz and Moesin may mediate the proper organization of actin in a local domain, which in turn stabilizes E-cadherin. These results provide a mechanism for the spatial order of actin organization underlying junction stabilization in primary embryonic epithelia.

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Figure 1: Epithelial integrity, adherens junctions stability and actin organization in embryos deficient for btsz or e-cad.
Figure 2: Btsz localization in the AJR requires Par-3 and PtdIns(4,5)P2.
Figure 3: Btsz interacts with Moesin.
Figure 4: Btsz and Moesin cooperate to stabilize E-cad.

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Acknowledgements

We thank all those who gave us reagents, in particular J. Serano and G. Rubin for fly strains and plasmids, A. Wodarz, A. Zelhof, R. Fehon, F. Payre, E. Suzuki, M. Peifer, H. Oda for flies, antibodies or plasmids; J. Großhans and R. Paro for reagents for synthesizing dsRNA probes for the RNAi screen. We also thank J. Knoblich for suggesting the PIPstrip experiment, members of our group for discussions, S. Kerridge and A. Lebivic for comments on the manuscript. This work was supported by the Association pour la Recherche contre le Cancer (ARC, subvention libre 5179), the CNRS, the Fondation pour la Recherche Médicale (FRM), the Fondation Schlumberger pour l'Education et la Recherche (FSER) and the EMBO Young Investigator Programme. F.P. was supported by the CNRS (bourse BDI) and by the Académie de médecine.

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  1. Institut de Biologie du Développement de Marseille Luminy (IBDML) UMR 6216, CNRS-Université de la Méditerrannée. Campus de Luminy, case 907, 13288, Marseille, cedex 09, France

    Fanny Pilot, Jean-Marc Philippe, Céline Lemmers & Thomas Lecuit

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Correspondence to Thomas Lecuit.

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Supplementary information

Supplementary Methods

This file contains Supplementary Methods and legends to Supplementary Figures and Supplementary Movies. (DOC 71 kb)

Supplementary Figure S1

Phenotypes in btszRNAi and btszK13-4/btszK13-4 mutant embryos. (PDF 663 kb)

Supplementary Figure S2

Phenotypes in btsz RNAi embryos, mutants and morphants. (PDF 256 kb)

Supplementary Figure S3

Iin situ hybridization with btsz probes. (PDF 317 kb)

Supplementary Figure S4

RT-PCR in btszRNAi embryos. (PDF 47 kb)

Supplementary Movie S1

Control RNAi embryo (control 1 probe). (MOV 1780 kb)

Supplementary Movie S2

btszRNAi (probe2) embryo with a strong phenotype (red category in Fig.S2a). (MOV 1618 kb)

Supplementary Movie S3

btszRNAi (probe2) embryo with a medium phenotype (orange category in Fig.S2a). (MOV 1553 kb)

Supplementary Movie S4

btszK13-4 mat-/-, zyg+/- mutant embryo without phenotype (blue category in Fig. S2f). (MOV 1506 kb)

Supplementary Movie S5

btszK13-4 mat-/-, zyg-/- mutant embryo with a strong phenotype (red category in Fig.S2f). (MOV 1332 kb)

Supplementary Movie S6

btszK13-4 mat-/-, zyg-/- mutant embryo with a medium phenotype (red category in Fig.S2f). (MOV 1537 kb)

Supplementary Movie S7

Confocal time lapse series showing E-cad-GFP at 15-s intervals in a btszRNAi embryo. (MOV 1308 kb)

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Pilot, F., Philippe, JM., Lemmers, C. et al. Spatial control of actin organization at adherens junctions by a synaptotagmin-like protein. Nature 442, 580–584 (2006). https://doi.org/10.1038/nature04935

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