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Abstract：
Ichthyosis with confetti (IWC) is a genodermatosis associated with dominant-negative variants in keratin 10 (KRT10) or keratin 1 (KRT1). These frameshift variants result in extended aberrant proteins, localized to the nucleus rather than the cytoplasm. This mislocalization is thought to occur as a result of the altered carboxy (C)-terminus, from poly-glycine to either a poly-arginine or -alanine tail. Previous studies on the type of C-terminus and subcellular localization of the respective mutant protein are divergent. In order to fully elucidate the pathomechanism of IWC, a greater understanding is critical. This study aimed to establish the consequences for localization and intermediate filament formation of altered keratin 10 (K10) C-termini. To achieve this, plasmids expressing distinct KRT10 variants were generated. Sequences encoded all possible reading frames of the K10 C-terminus as well as a nonsense variant. A keratinocyte line was transfected with these plasmids. Additionally, gene editing was utilized to introduce frameshift variants in exon 6 and exon 7 at the endogenous KRT10 locus. Cellular localization of aberrant K10 was observed via immunofluorescence using various antibodies. In each setting, immunofluorescence analysis demonstrated aberrant nuclear localization of K10 featuring an arginine-rich C-terminus. However, this was not observed with K10 featuring an alanine-rich C-terminus. Instead, the protein displayed cytoplasmic localization, consistent with wild-type and truncated forms of K10. This study demonstrates that, of the various 3\' frameshift variants of KRT10, exclusively arginine-rich C-termini lead to nuclear localization of K10.
摘要：
纸屑鱼鳞病（IWC）是与角蛋白10（KRT10）或角蛋白1（KRT1）中的显性阴性变体相关的遗传性皮肤病。这些移码变体导致延伸的异常蛋白，定位于细胞核而不是细胞质。这种错误定位被认为是由于羧基（C）末端的改变而发生的。从聚甘氨酸到聚精氨酸或丙氨酸尾。先前对相应突变蛋白的C末端类型和亚细胞定位的研究是不同的。为了充分阐明IWC的发病机制，更好的理解是至关重要的。这项研究旨在确定改变的角蛋白10（K10）C末端的定位和中间丝形成的后果。为了实现这一点，产生表达不同KRT10变体的质粒。序列编码K10C-末端的所有可能的阅读框以及无义变体。用这些质粒转染角质形成细胞系。此外,基因编辑用于在内源KRT10基因座的外显子6和外显子7中引入移码变体。使用各种抗体通过免疫荧光观察到异常K10的细胞定位。在每个设置中，免疫荧光分析表明K10的异常核定位具有富含精氨酸的C末端。然而,对于富含丙氨酸的C末端的K10没有观察到这种情况.相反，蛋白质显示细胞质定位，与K10的野生型和截短形式一致。这项研究表明,在KRT10的各种3'移码变体中，仅富含精氨酸的C末端导致K10的核定位。
