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Kazrin, a novel periplakin-interacting protein associated with desmosomes and the keratinocyte plasma membrane.

Groot KR, Sevilla LM, Nishi K, DiColandrea T, Watt FM - J. Cell Biol. (2004)

Bottom Line: Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes.On transfection, all three kazrin isoforms have similar subcellular distributions.We conclude that kazrin is a novel component of desmosomes that associates with periplakin.

View Article: PubMed Central - PubMed

Affiliation: Keratinocyte Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.

ABSTRACT
Periplakin forms part of the scaffold onto which the epidermal cornified envelope is assembled. The NH2-terminal 133 amino acids mediate association with the plasma membrane and bind a novel protein, kazrin. Kazrin is highly conserved and lacks homology to any known protein. There are four alternatively spliced transcripts, encoding three proteins with different NH2 termini. Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes. Kazrin colocalizes with periplakin and desmoplakin at desmosomes and with periplakin at the interdesmosomal plasma membrane, but its subcellular distribution is independent of periplakin. On transfection, all three kazrin isoforms have similar subcellular distributions. We conclude that kazrin is a novel component of desmosomes that associates with periplakin.

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Periplakin deletion analysis. (A) Schematic diagram of the periplakin constructs used for transient transfections. Domain nomenclature is described by Ruhrberg et al. (1997). All constructs had COOH-terminal HA tags. (B) Secondary structure predictions based on algorithms of Chou/Fasman and Garnier/Robson of the NH2-terminal 133 amino acids of periplakin. (C–K) Confocal projections of primary keratinocytes transfected with periplakin NH2-terminal constructs (C, P1/2N; D and E, P63; F–K, P133). (I–K) Cells were treated with Latrunculin B (200 ng/ml, 3 h) before fixation. Green, anti-HA; red, phalloidin-TRITC. C, E, H, and K show merged images. Bar: (C–H) 20 μm; (I–K) 10 μm.
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fig1: Periplakin deletion analysis. (A) Schematic diagram of the periplakin constructs used for transient transfections. Domain nomenclature is described by Ruhrberg et al. (1997). All constructs had COOH-terminal HA tags. (B) Secondary structure predictions based on algorithms of Chou/Fasman and Garnier/Robson of the NH2-terminal 133 amino acids of periplakin. (C–K) Confocal projections of primary keratinocytes transfected with periplakin NH2-terminal constructs (C, P1/2N; D and E, P63; F–K, P133). (I–K) Cells were treated with Latrunculin B (200 ng/ml, 3 h) before fixation. Green, anti-HA; red, phalloidin-TRITC. C, E, H, and K show merged images. Bar: (C–H) 20 μm; (I–K) 10 μm.

Mentions: The 495 NH2-terminal amino acids of periplakin (P1/2N), encoding the NN, Z, Y, and X subdomains, are sufficient for desmosomal and interdesmosomal plasma membrane localization (DiColandrea et al., 2000). At the interdesmosomal plasma membrane, P1/2N is concentrated at microvilli, defined here as actin-rich projections from the apical surface of cultured keratinocytes (DiColandrea et al., 2000; Fig. 1 C). Further deletion constructs were generated to precisely define the membrane interaction region (Fig. 1 A). Amino acids 1–133 of periplakin (P133), encoding the entire NN subdomain and the 16 amino acids that precede it, colocalized as efficiently as P1/2N (Fig. 1 C) with cortical actin (Fig. 1, F–H) and CD44 (not depicted) at microvilli. Addition of Latrunculin B caused redistribution of both P133 and P1/2N with actin (Fig. 1, I–K; DiColandrea et al., 2000).


Kazrin, a novel periplakin-interacting protein associated with desmosomes and the keratinocyte plasma membrane.

Groot KR, Sevilla LM, Nishi K, DiColandrea T, Watt FM - J. Cell Biol. (2004)

Periplakin deletion analysis. (A) Schematic diagram of the periplakin constructs used for transient transfections. Domain nomenclature is described by Ruhrberg et al. (1997). All constructs had COOH-terminal HA tags. (B) Secondary structure predictions based on algorithms of Chou/Fasman and Garnier/Robson of the NH2-terminal 133 amino acids of periplakin. (C–K) Confocal projections of primary keratinocytes transfected with periplakin NH2-terminal constructs (C, P1/2N; D and E, P63; F–K, P133). (I–K) Cells were treated with Latrunculin B (200 ng/ml, 3 h) before fixation. Green, anti-HA; red, phalloidin-TRITC. C, E, H, and K show merged images. Bar: (C–H) 20 μm; (I–K) 10 μm.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172441&req=5

fig1: Periplakin deletion analysis. (A) Schematic diagram of the periplakin constructs used for transient transfections. Domain nomenclature is described by Ruhrberg et al. (1997). All constructs had COOH-terminal HA tags. (B) Secondary structure predictions based on algorithms of Chou/Fasman and Garnier/Robson of the NH2-terminal 133 amino acids of periplakin. (C–K) Confocal projections of primary keratinocytes transfected with periplakin NH2-terminal constructs (C, P1/2N; D and E, P63; F–K, P133). (I–K) Cells were treated with Latrunculin B (200 ng/ml, 3 h) before fixation. Green, anti-HA; red, phalloidin-TRITC. C, E, H, and K show merged images. Bar: (C–H) 20 μm; (I–K) 10 μm.
Mentions: The 495 NH2-terminal amino acids of periplakin (P1/2N), encoding the NN, Z, Y, and X subdomains, are sufficient for desmosomal and interdesmosomal plasma membrane localization (DiColandrea et al., 2000). At the interdesmosomal plasma membrane, P1/2N is concentrated at microvilli, defined here as actin-rich projections from the apical surface of cultured keratinocytes (DiColandrea et al., 2000; Fig. 1 C). Further deletion constructs were generated to precisely define the membrane interaction region (Fig. 1 A). Amino acids 1–133 of periplakin (P133), encoding the entire NN subdomain and the 16 amino acids that precede it, colocalized as efficiently as P1/2N (Fig. 1 C) with cortical actin (Fig. 1, F–H) and CD44 (not depicted) at microvilli. Addition of Latrunculin B caused redistribution of both P133 and P1/2N with actin (Fig. 1, I–K; DiColandrea et al., 2000).

Bottom Line: Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes.On transfection, all three kazrin isoforms have similar subcellular distributions.We conclude that kazrin is a novel component of desmosomes that associates with periplakin.

View Article: PubMed Central - PubMed

Affiliation: Keratinocyte Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.

ABSTRACT
Periplakin forms part of the scaffold onto which the epidermal cornified envelope is assembled. The NH2-terminal 133 amino acids mediate association with the plasma membrane and bind a novel protein, kazrin. Kazrin is highly conserved and lacks homology to any known protein. There are four alternatively spliced transcripts, encoding three proteins with different NH2 termini. Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes. Kazrin colocalizes with periplakin and desmoplakin at desmosomes and with periplakin at the interdesmosomal plasma membrane, but its subcellular distribution is independent of periplakin. On transfection, all three kazrin isoforms have similar subcellular distributions. We conclude that kazrin is a novel component of desmosomes that associates with periplakin.

Show MeSH
Related in: MedlinePlus