Plakoglobin phosphorylation at serine 665 is capable of stabilizing cadherin-mediated adhesion in keratinocytes

TL;DR

Phosphorylation of plakoglobin at serine 665 helps stabilize skin cell adhesion in pemphigus, a disease where the skin blisters due to autoantibodies.

Contribution

This study shows that phosphorylation of plakoglobin at S665 is essential for apremilast's protective effects in pemphigus.

Findings

PG-S665A mice showed reduced keratin insertion and failed apremilast protection against blistering.

Phosphorylated PG at S665 colocalized with desmoplakin and keratin filaments at desmosomes.

Increased cAMP improved keratin insertion into desmosomal plaques in human skin.

Abstract

In pemphigus, autoantibodies against the desmosomal cadherins desmoglein (DSG) DSG1 and DSG3 cause intraepidermal blistering. Recently, we found that increasing cAMP with the phosphodiesterase-4 inhibitor apremilast stabilizes keratinocyte cohesion in pemphigus. This effect is paralleled by phosphorylation of the desmosomal plaque protein plakoglobin (PG) at serine 665 (S665). Here, we investigated the relevance of PG phosphorylation at S665 for stabilization of keratinocyte cohesion and further characterized the underlying mechanisms. Ultrastructural analysis of a recently established PG-S665 phospho-deficient mouse model (PG-S665A) showed diminished keratin insertion. Accordingly, the protective effect of apremilast against pemphigus autoantibody-induced skin blistering was diminished, and apremilast failed to restore alterations of the keratin cytoskeleton in PG-S665A mice. Keratinocytes derived from PG-S665A mice revealed a disorganized keratin cytoskeleton and reduced single-molecule binding strength of DSG3. In line with this, in ex vivo human skin, increased cAMP augmented keratin insertion into desmosomal plaques. Additionally, PG phosphorylated at S665 colocalized with desmoplakin and keratin filaments anchoring to desmosomes and increased cAMP-accelerated assembly of desmosomes. Taken together, phosphorylation of PG at S665 was crucial for protective effects of apremilast in pemphigus and for maintenance of DSG3 binding and keratin filament anchorage to desmosomes.

Phosphorylation of Pg at S665 is crucial for protective effects of apremilast in pemphigus and for maintenance of Dsg3 binding and keratin filament anchorage to desmosomes.

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