A Polybasic Domain in aPKC Mediates Par6-Dependent Control of Membrane Targeting and Kinase Activity

Wei, Dong; Liu, Juan; Zhang, Xintong; Wu, Yan; Lettieri, Kaela S.; Hammond, Gerald; Hong, Yang

doi:10.1101/588624

Cited by 13 publications

(27 citation statements)

References 64 publications

(95 reference statements)

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“…We also found that depletion of PAR-3 from the epidermis resulted in a partial loss of PAR-6. This is consistent with findings that multiple mechanisms can promote cortical recruitment of Par6-aPKC, including binding of Par6 to Par3 or Cdc42, and binding of aPKC to phospholipids (Dong et al, 2020;Hong et al, 2003;Hutterer et al, 2004;Joberty et al, 2000;Nagai-Tamai et al, 2002;Nunes de Almeida et al, 2019;Rodriguez et al, 2017;Wang et al, 2017;Wodarz et al, 2000). Conversely, the apical localization of PAR-3 was dependent upon PKC-3.…”

Section: Interdependencies Between the Par Proteinssupporting

confidence: 91%

Epidermal PAR-6 and PKC-3 are essential for postembryonic development ofCaenorhabditis elegansand control non-centrosomal microtubule organization

Castiglioni

Pires

Bertolini

et al. 2020

Preprint

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The cortical polarity regulators PAR-6, PKC-3 and PAR-3 are essential for the polarization of a broad variety of cell types in multicellular animals, from the first asymmetric division of the C. elegans zygote to apical–basal polarization of epithelial cells. In C. elegans, the roles of the PAR proteins in embryonic development have been extensively studied, yet little is known about their functions during larval development. Using auxin-inducible protein depletion, we here show that PAR-6 and PKC-3, but not PAR-3, are essential for postembryonic development. We also demonstrate that PAR-6 and PKC-3 are required in the epidermal epithelium to support animal growth and molting, and the proper timing and pattern of seam cell divisions. Finally, we uncovered a novel role for PAR-6 in controlling the organization of non-centrosomal microtubule arrays in the epidermis. PAR-6 was required for the localization of the microtubule organizer NOCA-1/Ninein, and microtubule defects in a noca-1 mutant are highly similar to those caused by epidermal PAR-6 depletion. As NOCA-1 physically interacts with PAR-6, we propose that PAR-6 promotes non-centrosomal microtubule organization through localization of NOCA-1/Ninein.SummaryUsing inducible protein degradation, we show that PAR-6 and PKC-3/aPKC are essential for postembryonic development of C. elegans and control the organization of non-centrosomal microtubule bundles in the epidermis, likely through recruitment of NOCA-1/Ninein.

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Section: Interdependencies Between the Par Proteinssupporting

confidence: 91%

Epidermal PAR-6 and PKC-3 are essential for postembryonic development ofCaenorhabditis elegansand control non-centrosomal microtubule organization

Castiglioni

Pires

Bertolini

et al. 2020

Preprint

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“…To further investigate the electrostatic PM targeting of Dlg in vivo, we used previously established hypoxia assays in which we acutely and reversibly deplete phospholipids PIP2 and PI4P in the PM (Dong et al, 2015) (data not shown). Similar to polybasic polarity proteins Lgl (Dong et al, 2015) and aPKC (Dong et al, 2020), in follicular cells hypoxia induced acute loss of PM Dlg::GFP which was reversed by subsequent reoxygenation (Fig. 1D).…”

Section: Resultsmentioning

confidence: 84%

“…While the electrostatic PM targeting of Lgl is now well established (Bailey and Prehoda, 2015;Dong et al, 2015), the exact mechanisms targeting Dlg to cell cortex or PM remain to be fully elucidated. Here we report that Dlg, like recently characterized polybasic polarity proteins Lgl and aPKC (Dong et al, 2020) contains a positively charged polybasic domain that targets Dlg to the PM and is necessary for Dlg to regulate polarity and tumorigenesis. Our results suggest that Scrib specifically enhances the electrostatic PM targeting of Dlg which is regulated by potential phosphorylation-dependent allosteric controls.…”

Section: Introductionmentioning

confidence: 79%

“…To investigate whether the PB domain is required for targeting Dlg to PM in vivo, we generated transgenic flies expressing Dlg::GFP, Dlg KR6Q ::GFP and Dlg KR15Q ::GFP under the ubiquitin promoter (Dong et al, 2020). Although dlg locus apparently expresses numerous isoforms, our Dlg::GFP based on isoform G fully rescued null mutants of dlg [A] (Haelterman et al, 2014;Ventura et al, 2020) and showed typical basolateral PM/cortex localization in follicular cells ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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Electrostatic Plasma Membrane Targeting Contributes to Dlg Function in Cell Polarity and Tumorigenesis

Liu

Wei

Yan

et al. 2020

Preprint

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SUMMARYDiscs large (Dlg) is an essential polarity protein and a tumor suppressor originally characterized in Drosophila but is also well conserved in vertebrates. Like the majority of polarity proteins, plasma membrane (PM)/cortical localization of Dlg is required for its function in regulating apical-basal polarity and tumorigenesis, but the exact mechanisms targeting Dlg to PM remain to be unclear. Here we show that, similar to recently discovered polybasic polarity proteins such as Lgl and aPKC, Dlg also contains a positively charged polybasic domain that electrostatically binds the PM phosphoinositides PI4P and PI(4,5)P2. Electrostatic targeting by the polybasic domain acts as the primary mechanism localizing Dlg to the PM in follicular and early embryonic epithelial cells, and is crucial for Dlg to regulate both polarity and tumorigenesis. The electrostatic PM targeting of Dlg is controlled by a potential phosphorylation-dependent allosteric regulation of its polybasic domain, and is specifically enhanced by interactions between Dlg and another basolateral polarity protein and tumor suppressor Scrib. Our studies highlight an increasingly significant role of electrostatic PM targeting of polarity proteins in regulating cell polarity.

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“…One mechanism that could localize Scrib to the cortex is a phospholipid-binding polybasic motif (PBM), as seen in other polarized proteins, including Lgl and aPKC (13,14,31). However, an obvious PBM is not seen in the Scrib protein sequence.…”

Section: Dlg Stabilizes Scrib At the Cortexmentioning

confidence: 99%

Distinct activities of Scrib module proteins organize epithelial polarity

Khoury

Bilder

2019

Preprint

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A polarized architecture is central to both epithelial structure and function. In many cells, polarity involves mutual antagonism between the Par complex and the Scrib module. While molecular mechanisms underlying Par-mediated apical determination are well-understood, how Scrib module proteins specify the basolateral domain remains unknown. Here, we demonstrate dependent and independent activities of Scrib, Dlg and Lgl using the Drosophila follicle epithelium. Our data support a linear hierarchy for localization, but rule out previously proposed protein-protein interactions as essential for polarization. Membrane recruitment of Scrib does not require palmitoylation or polar phospholipid binding but instead an independent cortically-stabilizing activity of Dlg. Scrib and Dlg do not directly antagonize aPKC, but may instead restrict aPKC localization by enabling the aPKC-inhibiting activity of Lgl. Importantly, while Scrib, Dlg and Lgl are each required, all three together are not sufficient to antagonize the Par complex. Our data demonstrate previously unappreciated diversity of function within the Scrib module and begin to define the elusive molecular functions of Scrib and Dlg. SIGNIFICANCE STATEMENTTo enable their physiological functions, cells must polarize their plasma membrane. In many epithelia, polarity is regulated by balanced activity of the apical Par complex and basolateral Scribble module. While the former is understood in molecular detail, little is known about how the latter works. We identify distinct functions of the three Scribble module proteins, separating independent roles in a localization hierarchy from cooperative roles in apical polarity antagonism and showing that they are not together sufficient to specify basolateral identity. This work establishes an essential basis for a mechanistic understanding of this core polarity machinery that controls processes ranging from stem cell divisions to organ morphogenesis across animal species.

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A Polybasic Domain in aPKC Mediates Par6-Dependent Control of Membrane Targeting and Kinase Activity

Cited by 13 publications

References 64 publications

Epidermal PAR-6 and PKC-3 are essential for postembryonic development ofCaenorhabditis elegansand control non-centrosomal microtubule organization

Epidermal PAR-6 and PKC-3 are essential for postembryonic development ofCaenorhabditis elegansand control non-centrosomal microtubule organization

Electrostatic Plasma Membrane Targeting Contributes to Dlg Function in Cell Polarity and Tumorigenesis

Distinct activities of Scrib module proteins organize epithelial polarity

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