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Dynamics of core planar polarity protein turnover and stable assembly into discrete membrane subdomains.

Strutt H, Warrington SJ, Strutt D - Dev. Cell (2011)

Bottom Line: The core planar polarity proteins localize asymmetrically to the adherens junctions of epithelial cells, where they have been hypothesized to assemble into intercellular complexes.Here, we show that the core proteins are preferentially distributed to discrete membrane subdomains ("puncta"), where they form asymmetric contacts between neighboring cells.Thus, we define the distinct roles of specific core proteins in the formation of asymmetric contacts between cells, which is a key event in the generation of coordinated cellular asymmetry.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Developmental and Biomedical Genetics, and Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.

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Asymmetric Localization of Core Proteins and Fmi Internalization in Prepupal Wings(A) Diagram of core protein distributions in inferred asymmetric intercellular complex. (B) Organization of individual complexes into domains of common polarity. (C–F) Prepupal wings. (C) Wild-type wing stained for Fmi. (D) fmiE59 clone marked by loss of β-gal (red), stained for extracellular Fmi (green), showing that apical nonjunctional staining is specific. (E and F) fz-EYFP (E) and stbm-EYFP (F) mosaics, stained for GFP (green) and Fmi (red). Arrows show localization of Fz-EYFP toward (E) or Stbm-EYFP away from (F) the wing margin. (G–J) Fmi antibody internalization with chase times up to 30 min in wild-type prepupal wings. (G) Quantitation of extracellular Fmi staining. For each time point n > 100 in 19 experiments. Here and in later figures, error bars are standard error of the mean, and asterisks indicate p values (p∗ ≤ 0.05; p∗∗ ≤ 0.01; p∗∗∗ ≤ 0.001; NS, not significant p > 0.05). Here, p values are relative to the previous time point. (H) Extracellular Fmi staining in apical XY sections. Note that apical staining at 0 min (yellow arrow) is reduced at later times, whereas junctional puncta become more distinct (white arrows). (I) Total Fmi staining in subapical XY sections. Arrows indicate intracellular Fmi puncta. (J) XZ sections of extracellular (green) and total (red) Fmi staining, showing apical junctional puncta (white arrows) and intracellular Fmi puncta (red arrows). Scale bars, 10 μm (C and D) or 2.5 μm (E, F, and H–J). See also Figures S1 and S2.
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fig1: Asymmetric Localization of Core Proteins and Fmi Internalization in Prepupal Wings(A) Diagram of core protein distributions in inferred asymmetric intercellular complex. (B) Organization of individual complexes into domains of common polarity. (C–F) Prepupal wings. (C) Wild-type wing stained for Fmi. (D) fmiE59 clone marked by loss of β-gal (red), stained for extracellular Fmi (green), showing that apical nonjunctional staining is specific. (E and F) fz-EYFP (E) and stbm-EYFP (F) mosaics, stained for GFP (green) and Fmi (red). Arrows show localization of Fz-EYFP toward (E) or Stbm-EYFP away from (F) the wing margin. (G–J) Fmi antibody internalization with chase times up to 30 min in wild-type prepupal wings. (G) Quantitation of extracellular Fmi staining. For each time point n > 100 in 19 experiments. Here and in later figures, error bars are standard error of the mean, and asterisks indicate p values (p∗ ≤ 0.05; p∗∗ ≤ 0.01; p∗∗∗ ≤ 0.001; NS, not significant p > 0.05). Here, p values are relative to the previous time point. (H) Extracellular Fmi staining in apical XY sections. Note that apical staining at 0 min (yellow arrow) is reduced at later times, whereas junctional puncta become more distinct (white arrows). (I) Total Fmi staining in subapical XY sections. Arrows indicate intracellular Fmi puncta. (J) XZ sections of extracellular (green) and total (red) Fmi staining, showing apical junctional puncta (white arrows) and intracellular Fmi puncta (red arrows). Scale bars, 10 μm (C and D) or 2.5 μm (E, F, and H–J). See also Figures S1 and S2.

Mentions: In both flies and vertebrates, the core proteins localize asymmetrically within cells prior to morphological signs of polarization (Rida and Chen, 2009; Roszko et al., 2009; Strutt and Strutt, 2009; Hashimoto and Hamada, 2010). In the Drosophila early pupal wing, asymmetric localization is seen within the adherens junction region, oriented toward the wing margin (Classen et al., 2005; Aigouy et al., 2010). Cell rearrangements then lead to a remodeling of asymmetry, such that it aligns on the proximodistal (PD) axis (Aigouy et al., 2010). Consequently, at the time that trichomes emerge, the seven-pass transmembrane protein Frizzled (Fz) localizes distally, together with the cytoplasmic proteins Dishevelled (Dsh) and Diego (Dgo), whereas the transmembrane protein Strabismus (Stbm, also known as Van Gogh) and the cytoplasmic protein Prickle (Pk) localize proximally, and the seven-pass transmembrane cadherin Flamingo (Fmi, also known as Starry Night) localizes both proximally and distally (Figure 1A; reviewed in Strutt and Strutt, 2009).


Dynamics of core planar polarity protein turnover and stable assembly into discrete membrane subdomains.

Strutt H, Warrington SJ, Strutt D - Dev. Cell (2011)

Asymmetric Localization of Core Proteins and Fmi Internalization in Prepupal Wings(A) Diagram of core protein distributions in inferred asymmetric intercellular complex. (B) Organization of individual complexes into domains of common polarity. (C–F) Prepupal wings. (C) Wild-type wing stained for Fmi. (D) fmiE59 clone marked by loss of β-gal (red), stained for extracellular Fmi (green), showing that apical nonjunctional staining is specific. (E and F) fz-EYFP (E) and stbm-EYFP (F) mosaics, stained for GFP (green) and Fmi (red). Arrows show localization of Fz-EYFP toward (E) or Stbm-EYFP away from (F) the wing margin. (G–J) Fmi antibody internalization with chase times up to 30 min in wild-type prepupal wings. (G) Quantitation of extracellular Fmi staining. For each time point n > 100 in 19 experiments. Here and in later figures, error bars are standard error of the mean, and asterisks indicate p values (p∗ ≤ 0.05; p∗∗ ≤ 0.01; p∗∗∗ ≤ 0.001; NS, not significant p > 0.05). Here, p values are relative to the previous time point. (H) Extracellular Fmi staining in apical XY sections. Note that apical staining at 0 min (yellow arrow) is reduced at later times, whereas junctional puncta become more distinct (white arrows). (I) Total Fmi staining in subapical XY sections. Arrows indicate intracellular Fmi puncta. (J) XZ sections of extracellular (green) and total (red) Fmi staining, showing apical junctional puncta (white arrows) and intracellular Fmi puncta (red arrows). Scale bars, 10 μm (C and D) or 2.5 μm (E, F, and H–J). See also Figures S1 and S2.
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fig1: Asymmetric Localization of Core Proteins and Fmi Internalization in Prepupal Wings(A) Diagram of core protein distributions in inferred asymmetric intercellular complex. (B) Organization of individual complexes into domains of common polarity. (C–F) Prepupal wings. (C) Wild-type wing stained for Fmi. (D) fmiE59 clone marked by loss of β-gal (red), stained for extracellular Fmi (green), showing that apical nonjunctional staining is specific. (E and F) fz-EYFP (E) and stbm-EYFP (F) mosaics, stained for GFP (green) and Fmi (red). Arrows show localization of Fz-EYFP toward (E) or Stbm-EYFP away from (F) the wing margin. (G–J) Fmi antibody internalization with chase times up to 30 min in wild-type prepupal wings. (G) Quantitation of extracellular Fmi staining. For each time point n > 100 in 19 experiments. Here and in later figures, error bars are standard error of the mean, and asterisks indicate p values (p∗ ≤ 0.05; p∗∗ ≤ 0.01; p∗∗∗ ≤ 0.001; NS, not significant p > 0.05). Here, p values are relative to the previous time point. (H) Extracellular Fmi staining in apical XY sections. Note that apical staining at 0 min (yellow arrow) is reduced at later times, whereas junctional puncta become more distinct (white arrows). (I) Total Fmi staining in subapical XY sections. Arrows indicate intracellular Fmi puncta. (J) XZ sections of extracellular (green) and total (red) Fmi staining, showing apical junctional puncta (white arrows) and intracellular Fmi puncta (red arrows). Scale bars, 10 μm (C and D) or 2.5 μm (E, F, and H–J). See also Figures S1 and S2.
Mentions: In both flies and vertebrates, the core proteins localize asymmetrically within cells prior to morphological signs of polarization (Rida and Chen, 2009; Roszko et al., 2009; Strutt and Strutt, 2009; Hashimoto and Hamada, 2010). In the Drosophila early pupal wing, asymmetric localization is seen within the adherens junction region, oriented toward the wing margin (Classen et al., 2005; Aigouy et al., 2010). Cell rearrangements then lead to a remodeling of asymmetry, such that it aligns on the proximodistal (PD) axis (Aigouy et al., 2010). Consequently, at the time that trichomes emerge, the seven-pass transmembrane protein Frizzled (Fz) localizes distally, together with the cytoplasmic proteins Dishevelled (Dsh) and Diego (Dgo), whereas the transmembrane protein Strabismus (Stbm, also known as Van Gogh) and the cytoplasmic protein Prickle (Pk) localize proximally, and the seven-pass transmembrane cadherin Flamingo (Fmi, also known as Starry Night) localizes both proximally and distally (Figure 1A; reviewed in Strutt and Strutt, 2009).

Bottom Line: The core planar polarity proteins localize asymmetrically to the adherens junctions of epithelial cells, where they have been hypothesized to assemble into intercellular complexes.Here, we show that the core proteins are preferentially distributed to discrete membrane subdomains ("puncta"), where they form asymmetric contacts between neighboring cells.Thus, we define the distinct roles of specific core proteins in the formation of asymmetric contacts between cells, which is a key event in the generation of coordinated cellular asymmetry.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Developmental and Biomedical Genetics, and Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.

Show MeSH
Related in: MedlinePlus