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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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Kazrin localization in cultured keratinocytes. Human keratinocytes (A–L) and keratinocytes from wild-type (M and N) or periplakin  (O and P) mice were immunolabeled with antibodies to kazrin (K; A–J and M–P, green) and periplakin (PPL; A–D, red; and K and L, green), desmoplakin (DP; E–H and M–P, red) or keratins (IF; I–L, red). Boxes in A, E, I, K, M, and O are enlarged in B–D, F–H, J, L, N, and P, respectively. Bar: (A, E, I, K, M, and O) 20 μm; (B–D and F–H) 7 μm; (J and L) 5.4 μm; (N and P) 5 μm. (Q–U) Kazrin isoforms (Q, kazrin A; R, kazrin B; S–U, kazrin C) with NH2-terminal HA tags were transiently transfected into primary human keratinocytes. (Q–U) green, anti-HA; red, phalloidin-TRITC. Q–S are composite images of z-stacks. Arrowheads in S indicate position of orthogonal view shown in T. Area boxed in S is shown as single confocal slice in U. Bar: (Q–S) 20 μm; (T) 11.5 μm; (U) 6 μm.
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fig5: Kazrin localization in cultured keratinocytes. Human keratinocytes (A–L) and keratinocytes from wild-type (M and N) or periplakin (O and P) mice were immunolabeled with antibodies to kazrin (K; A–J and M–P, green) and periplakin (PPL; A–D, red; and K and L, green), desmoplakin (DP; E–H and M–P, red) or keratins (IF; I–L, red). Boxes in A, E, I, K, M, and O are enlarged in B–D, F–H, J, L, N, and P, respectively. Bar: (A, E, I, K, M, and O) 20 μm; (B–D and F–H) 7 μm; (J and L) 5.4 μm; (N and P) 5 μm. (Q–U) Kazrin isoforms (Q, kazrin A; R, kazrin B; S–U, kazrin C) with NH2-terminal HA tags were transiently transfected into primary human keratinocytes. (Q–U) green, anti-HA; red, phalloidin-TRITC. Q–S are composite images of z-stacks. Arrowheads in S indicate position of orthogonal view shown in T. Area boxed in S is shown as single confocal slice in U. Bar: (Q–S) 20 μm; (T) 11.5 μm; (U) 6 μm.

Mentions: In differentiating primary human keratinocytes, kazrin was observed at cell–cell borders and, to a variable degree, in the nucleus (Fig. 5, A–L). In stratified cultures, large numbers of desmosomes assemble at the interface between the apical surfaces of basal layer cells and the basal surfaces of suprabasal cells. Kazrin partially colocalized with desmoplakin and periplakin at the desmosomes and with periplakin at the interdesmosomal plasma membrane (Fig. 5, A–H). Kazrin was localized to puncta, corresponding to desmosomes, where IFs in neighboring cells aligned (Fig. 5, I–L). The subcellular distribution of kazrin was indistinguishable in keratinocyte lines generated from periplakin- and wild-type mice (Fig. 5, M–P), demonstrating that kazrin was not dependent on periplakin for its localization. The shift in kazrin localization to cell–cell borders in differentiating periplakin- keratinocytes suggests an association with additional desmosomal components that are up-regulated during terminal differentiation.


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)

Kazrin localization in cultured keratinocytes. Human keratinocytes (A–L) and keratinocytes from wild-type (M and N) or periplakin  (O and P) mice were immunolabeled with antibodies to kazrin (K; A–J and M–P, green) and periplakin (PPL; A–D, red; and K and L, green), desmoplakin (DP; E–H and M–P, red) or keratins (IF; I–L, red). Boxes in A, E, I, K, M, and O are enlarged in B–D, F–H, J, L, N, and P, respectively. Bar: (A, E, I, K, M, and O) 20 μm; (B–D and F–H) 7 μm; (J and L) 5.4 μm; (N and P) 5 μm. (Q–U) Kazrin isoforms (Q, kazrin A; R, kazrin B; S–U, kazrin C) with NH2-terminal HA tags were transiently transfected into primary human keratinocytes. (Q–U) green, anti-HA; red, phalloidin-TRITC. Q–S are composite images of z-stacks. Arrowheads in S indicate position of orthogonal view shown in T. Area boxed in S is shown as single confocal slice in U. Bar: (Q–S) 20 μm; (T) 11.5 μm; (U) 6 μm.
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fig5: Kazrin localization in cultured keratinocytes. Human keratinocytes (A–L) and keratinocytes from wild-type (M and N) or periplakin (O and P) mice were immunolabeled with antibodies to kazrin (K; A–J and M–P, green) and periplakin (PPL; A–D, red; and K and L, green), desmoplakin (DP; E–H and M–P, red) or keratins (IF; I–L, red). Boxes in A, E, I, K, M, and O are enlarged in B–D, F–H, J, L, N, and P, respectively. Bar: (A, E, I, K, M, and O) 20 μm; (B–D and F–H) 7 μm; (J and L) 5.4 μm; (N and P) 5 μm. (Q–U) Kazrin isoforms (Q, kazrin A; R, kazrin B; S–U, kazrin C) with NH2-terminal HA tags were transiently transfected into primary human keratinocytes. (Q–U) green, anti-HA; red, phalloidin-TRITC. Q–S are composite images of z-stacks. Arrowheads in S indicate position of orthogonal view shown in T. Area boxed in S is shown as single confocal slice in U. Bar: (Q–S) 20 μm; (T) 11.5 μm; (U) 6 μm.
Mentions: In differentiating primary human keratinocytes, kazrin was observed at cell–cell borders and, to a variable degree, in the nucleus (Fig. 5, A–L). In stratified cultures, large numbers of desmosomes assemble at the interface between the apical surfaces of basal layer cells and the basal surfaces of suprabasal cells. Kazrin partially colocalized with desmoplakin and periplakin at the desmosomes and with periplakin at the interdesmosomal plasma membrane (Fig. 5, A–H). Kazrin was localized to puncta, corresponding to desmosomes, where IFs in neighboring cells aligned (Fig. 5, I–L). The subcellular distribution of kazrin was indistinguishable in keratinocyte lines generated from periplakin- and wild-type mice (Fig. 5, M–P), demonstrating that kazrin was not dependent on periplakin for its localization. The shift in kazrin localization to cell–cell borders in differentiating periplakin- keratinocytes suggests an association with additional desmosomal components that are up-regulated during terminal differentiation.

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