A conserved polybasic domain mediates plasma membrane targeting of Lgl and its regulation by hypoxia

Wei, Dong; Zhang, Xuejing; Liu, Weijie; Chen, Yi‐Jiun; Huang, Juan; Austin, Erin; Celotto, Alicia M.; Jiang, Wendy Z.; Palladino, Michael J.; Jiang, Yu; Hammond, Gerald; Hong, Yang

doi:10.1083/jcb.201503067

Cited by 73 publications

(161 citation statements)

References 46 publications

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“…Drosophila stocks: Knock-out alleles of lgl GX ("lgl KO ") and crb GX ("crb KO ") and knock-in alleles of lgl::GFP and lgl S5A ::GFP were previously described (Dong et al, 2015;Huang et al, 2009). w; αTub67C-Gal4 V2H ; αTub67C-Gal4 V37 ("Mat-Gal4", a gift from Dr. Mark Peifer, University of North Carolina at Chapel Hill, USA) (Schaefer et al, 2018).…”

Section: Fly Stocks and Geneticsmentioning

confidence: 99%

“…Liposome pull-down assays. Bacteria BL21 was used to express and purify GST and GST fusion proteins as previously described (Dong et al, 2015). Liposomal binding assays were carried out as described (Kim et al, 2008).…”

Section: Molecular Cloningmentioning

confidence: 99%

“…Moreover, polybasic domains in Lgl, Numb and Miranda also contain conserved serine residues that can be phosphorylated by another key polarity protein atypical PKC (aPKC) (Bailey and Prehoda, 2015;Dong et al, 2015). Similar to the phosphorylation of the polybasic ED domain in MARCKS by PKC (Arbuzova et al, 2002), aPKC-phosphorylation neutralizes the positive charges in a polybasic domain therefore inhibits its electrostatic binding to phospholipids on the PM (Bailey and Prehoda, 2015;Dong et al, 2015). Such aPKC-dependent inhibition serves an elegant mechanism to polarize the PM targeting of polybasic polarity proteins, allowing apically localized aPKC to limit Lgl, Numb and Miranda to basolateral PM.…”

Section: Introductionmentioning

confidence: 99%

“…Polybasic domains can bind to PM specifically, as the inner surface of PM is the most negatively charged membrane surface due to membrane phosphatidylserine (Yeung et al, 2008) and its unique enrichment of polyphosphoinositides PI4P and PI(4,5)P2 (PIP2) . Moreover, polybasic domains in Lgl, Numb and Miranda also contain conserved serine residues that can be phosphorylated by another key polarity protein atypical PKC (aPKC) (Bailey and Prehoda, 2015;Dong et al, 2015). Similar to the phosphorylation of the polybasic ED domain in MARCKS by PKC (Arbuzova et al, 2002), aPKC-phosphorylation neutralizes the positive charges in a polybasic domain therefore inhibits its electrostatic binding to phospholipids on the PM (Bailey and Prehoda, 2015;Dong et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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

Wei

Liu

Zhang

et al. 2019

Preprint

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Mechanisms coupling the atypical PKC (aPKC) kinase activity to its subcellular localization are essential for regulating cell polarization but remain to be fully elucidated. Unlike other members of the PKC family, aPKC has no well-defined or calcium binding domains, leading to the assumption that its subcellular localization relies exclusively on protein-protein interactions. Here we show that in both Drosophila and mammalian cells the pseudosubstrate region (PSr) of aPKC acts as a polybasic domain sufficient to target aPKC to the plasma membrane (PM) via electrostatic binding to PM phosphoinositides. PM-targeting of aPKC requires the physical interaction between the N-terminal PB1 domains in both aPKC and Par-6, which not only allosterically exposes PSr to PM-binding, but also inhibits aPKC kinase activity.Such kinase inhibition requires the C-terminal domain of Par-6 and can be relieved through Par-6 interaction with apical polarity protein Crumbs. Our data suggest a mechanism in which allosteric regulation of polybasic PSr in aPKC by Par-6 couples the control of both aPKC subcellular localization and spatial activation of its kinase activity.3

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Section: Fly Stocks and Geneticsmentioning

confidence: 99%

Section: Molecular Cloningmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Wei

Liu

Zhang

et al. 2019

Preprint

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“…These substrates can localize to cortical regions that lack the Par complex but are removed from the cortex once they enter the Par domain (Figure 1B). For at least several aPKC substrates, this mechanism appears to involve phosphorylation of short motifs enriched for basic and hydrophobic residues that directly interact with phospholipids 26,27 . Phosphorylation of the motif alters its electrostatic character thereby reducing the affinity for the membrane and causing displacement of the substrate into the cytoplasm.…”

Section: Cellular Mechanisms Of Fate Determination By Apkcmentioning

confidence: 99%

Molecular Control of Atypical Protein Kinase C: Tipping the Balance between Self-Renewal and Differentiation

Drummond

Prehoda

2016

Journal of Molecular Biology

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Complex organisms are faced with the challenge of generating and maintaining diverse cell types, ranging from simple epithelia, to neurons and motile immune cells1–3. To meet this challenge, a complex set of regulatory pathways controls nearly every aspect of cell growth and function, including genetic and epigenetic programming, cytoskeleton dynamics, and protein trafficking. The far reach of cell fate specification pathways makes it particularly catastrophic when they malfunction, both during development and for tissue homeostasis in adult organisms. Furthermore, the promise of stem cells as a therapeutic derives from their ability to deftly navigate the multitude of pathways that control cell fate4. How the molecular components that make up these pathways function to specify cell fate is beginning to become clear. Work from diverse systems suggests that the atypical Protein Kinase C (aPKC) is a key regulator of cell fate decisions in metazoans5–7. Here we examine some of the diverse physiological outcomes of aPKC’s function in differentiation, along with the molecular pathways that control aPKC, and those that are responsive to changes in its catalytic activity.

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PAR3–PAR6–atypical PKC polarity complex proteins in neuronal polarization

Hapak

Rothlin

Ghosh

2018

Cell. Mol. Life Sci.

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Polarity is a fundamental feature of cells. Protein complexes, including the PAR3-PAR6-aPKC complex, have conserved roles in establishing polarity across a number of eukaryotic cell types. In neurons, polarity is evident as distinct axonal versus dendritic domains. The PAR3, PAR6, and aPKC proteins also play important roles in neuronal polarization. During this process, either aPKC kinase activity, the assembly of the PAR3-PAR6-aPKC complex or the localization of these proteins is regulated downstream of a number of signaling pathways. In turn, the PAR3, PAR6, and aPKC proteins control various effector molecules to establish neuronal polarity. Herein, we discuss the many signaling mechanisms and effector functions that have been linked to PAR3, PAR6, and aPKC during the establishment of neuronal polarity.

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A conserved polybasic domain mediates plasma membrane targeting of Lgl and its regulation by hypoxia

Abstract: The plasma membrane targeting of Lgl, a key polarity and tumor suppressor protein, is mediated by electrostatic interactions between a polybasic motif in Lgl and phospholipids on the plasma membrane, and this mechanism is regulated by hypoxia and aPKC-phosphorylation.

Cited by 73 publications

References 46 publications

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

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

Molecular Control of Atypical Protein Kinase C: Tipping the Balance between Self-Renewal and Differentiation

PAR3–PAR6–atypical PKC polarity complex proteins in neuronal polarization

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