title: The DSPII splice variant is crucial for desmosome-mediated adhesion in HaCaT keratinocytes.
abstract: Desmosomes are intercellular junctions specialised for strong adhesion that are prominent in the epidermis and heart muscle. Defective desmosomal function due to inherited mutations in the constitutive desmosomal gene desmoplakin (DSP) causes skin or heart disorders and in some instances both. Different mutations have different disease-causing molecular mechanisms as evidenced by the varying phenotypes resulting from mutations affecting different domains of the same protein, but the majority of these mechanisms remain to be determined. Here, we studied two mutations in DSP that lead to different dosages of the two major DSP splice variants, DSPI and DSPII, and compared their molecular mechanisms. One of the mutations results in total DSP haploinsufficiency and is associated with autosomal dominant striate palmoplantar keratoderma (PPK). The other leads to complete absence of DSPI and the minor isoform DSPIa but normal levels of DSPII, and is associated with autosomal recessive epidermolytic PPK, woolly hair and severe arrhythmogenic dilated cardiomyopathy. Using siRNA treatments to mimic these two mutations and additionally a DSPII-specific siRNA, we found striking differences between DSP isoforms with respect to keratinocyte adhesion upon cellular stress with DSPII being the key component in intermediate filament (IF) stability and desmosome-mediated adhesion. In addition, reduction in DSP expression reduced the amount of plakophilin 1, desmocollin (DSC) 2 and DSC3 with DSPI having a greater influence than DSPII on the expression levels of DSC3. These results suggest that the two major DSP splice variants are not completely redundant in function and that DSPII dosage is particularly important for desmosomal adhesion in the skin.
authors.affiliation: Centre for Cutaneous Research, The Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, 4 Newark Street, London E1 2AT,  UK.
authors.forename: Rita M
authors.initials: RM
authors.lastname: Cabral
authors.forename: Daniel
authors.initials: D
authors.lastname: Tattersall
authors.forename: Vishal
authors.initials: V
authors.lastname: Patel
authors.forename: Graham D
authors.initials: GD
authors.lastname: McPhail
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Hatzimasoura
authors.forename: Dominic J
authors.initials: DJ
authors.lastname: Abrams
authors.forename: Andrew P
authors.initials: AP
authors.lastname: South
authors.forename: David P
authors.initials: DP
authors.lastname: Kelsell
mesh_terms: D002448:Cell Adhesion; D002460:Cell Line; D051180:Desmoplakins / Q000235:genetics* / Q000378:metabolism*; D003896:Desmosomes / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing
chemical_list: D051180:Desmoplakins; D020033:Protein Isoforms