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The Scrib module proteins, Scrib, Dlg, and Lgl, are conserved regulators of cell polarity in diverse biological contexts. Originally discovered as neoplastic tumor suppressors in the fruit fly Drosophila melanogaster, disruption of Scrib module components leads to tumorigenesis in mammalian epithelia and is associated with human cancers. With multiple protein interacting domains, Scrib module proteins function as determinants of basolateral identity in epithelial cells with apical-basal polarity while acting as signaling platform scaffold proteins. Recent studies have further revealed novel roles of the Scrib module in the control of epithelial architecture, ranging from polarity establishment and tricellular junction formation to planar spindle orientation during cell division. This review updates the current understanding of the molecular nature and physiological functions of the Scrib module with a focus on in vivo studies, providing a framework for how these protein dynamics affect the processes of epithelial organization., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The Drosophila melanogaster wing imaginal disc is an epithelial sac that exhibits dramatic tissue growth during the larval stage. With its simple morphology and accessibility of genetic tools, studies using the wing disc have contributed to the understanding of the mechanisms of epithelial homeostasis including the control of mitotic spindle orientation. This chapter describes a detailed protocol for analyzing epithelial architecture and planar orientation of the mitotic spindle in the wing disc epithelium. The rapid dissection method, effective immunostaining, and mounting tips described here facilitate genetic and cell biological studies of the wing disc and can be applied to a wide array of studies using various Drosophila tissues.

Proper orientation of the mitotic spindle is essential for cell fate determination, tissue morphogenesis, and homeostasis. During epithelial proliferation, planar spindle alignment ensures the maintenance of polarized tissue architecture, and aberrant spindle orientation can disrupt epithelial integrity. Nevertheless, in vivo mechanisms that restrict the mitotic spindle to the plane of the epithelium remain poorly understood. Here we show that the junction-localized tumor suppressors Scribbled (Scrib) and Discs large (Dlg) control planar spindle orientation via Mud and 14-3-3 proteins in the Drosophila wing disc epithelium. During mitosis, Scrib is required for the junctional localization of Dlg, and both affect mitotic spindle movements. Using coimmunoprecipitation and mass spectrometry, we identify 14-3-3 proteins as Dlg-interacting partners and further report that loss of 14-3-3s causes both abnormal spindle orientation and disruption of epithelial architecture as a consequence of basal cell delamination and apoptosis. Combined, these biochemical and genetic analyses indicate that 14-3-3s function together with Scrib, Dlg, and Mud during planar cell division., (© 2019 Nakajima et al.)

Polarized epithelia are a foundation of organ and appendage structures throughout Metazoa and serve as a physical barrier to preserve physiological functions. In proliferating epithelia, planar cell division occurs by orienting the mitotic spindle within the plane of the epithelium to ensure tissue organization. Conversely, loss of tissue architecture is a hallmark of carcinoma, and aberrant spindle orientation is hypothesized to contribute to tissue disorganization through dysplasia and cell dissemination. Recent in vivo studies have uncovered a role of planar spindle alignment in the robust maintenance of tissue architecture, which accompanies homeostatic mechanisms such as cell delamination and re-integration of misplaced cells following abnormal cell division. Furthermore, perpendicular spindle orientation shifts have been suggested as causes of cell fate change and epithelial plasticity manifested by epithelial-to-mesenchymal transition. This review describes the mechanism by which planar spindle orientation is tightly regulated and discusses the roles of mitotic spindle orientation in epithelial development and disease.

During epithelial cell proliferation, planar alignment of the mitotic spindle coordinates the local process of symmetric cell cleavage with the global maintenance of polarized tissue architecture. Although the disruption of planar spindle alignment is proposed to cause epithelial to mesenchymal transition and cancer, the in vivo mechanisms regulating mitotic spindle orientation remain elusive. Here we demonstrate that the actomyosin cortex and the junction-localized neoplastic tumour suppressors Scribbled and Discs large 1 have essential roles in planar spindle alignment and thus the control of epithelial integrity in the Drosophila imaginal disc. We show that defective alignment of the mitotic spindle correlates with cell delamination and apoptotic death, and that blocking the death of misaligned cells is sufficient to drive the formation of basally localized tumour-like masses. These findings indicate a key role for junction-mediated spindle alignment in the maintenance of epithelial integrity, and also reveal a previously unknown cell-death-mediated tumour-suppressor function inherent in the polarized architecture of epithelia.