Molecular cloning and characterization of alternatively spliced mRNA isoforms from psoriatic skin encoding a novel member of the S100 family
Molecular cloning and characterization of alternatively spliced mRNA isoforms from psoriatic skin encoding a novel member of the S100 family
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DOI：
10.1096/fj.03-0148fje
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发表时间：
2003-08-01
期刊：
影响因子：
4.8
通讯作者：
Ruzicka, T
中科院分区：
文献类型：
--
作者：
Wolf, R;Mirmohammadsadegh, A;Ruzicka, T
为了鉴定银屑病相关基因，我们使用差异显示RT-PCR技术比较了正常皮肤和银屑病皮肤的基因表达。对病变皮肤中高度上调的650 bp cDNA片段（克隆110）进行序列分析，发现其与非编码cDNA （NICE-2）同源。通过随后的cDNA克隆，使用来自银屑病皮肤的RNA，我们已经鉴定出两个选择性剪接的mrna -异构体（0.5和4.4 kb），它们的非翻译区域的组成不同。通过序列比较，我们将这个名为S100A15的新基因定位到表皮分化复合体（染色体1q21）内的S100基因簇上。对推导出的氨基酸序列进行分析，发现该蛋白含有101个氨基酸，包含两个与S100A7具有高度同源性的潜在EF-hand基序。Northern blot杂交和半定量RT-PCR分析证实了S100A15在银屑病中的过表达，显示出不同水平的S100A15 mRNA亚型的表达。S100A15的原位杂交显示，与健康组织相比，银屑病皮肤的基底和基底上表皮层的染色明显增加。我们的数据表明，新的S100A15参与表皮分化和炎症，因此可能对牛皮癣和其他疾病的发病机制很重要。
In an effort to identify psoriasis-associated genes, we compared gene expression in normal and psoriatic skin, using differential display RT-PCR technique. Sequence analysis of a 650-bp cDNA fragment ( clone 110) that was highly up-regulated in lesional skin revealed homology to a noncoding cDNA (NICE-2). By subsequent cDNA cloning, using RNA from psoriatic skin, we have identified two alternatively spliced mRNA-isoforms (0.5 and 4.4 kb), which differ in composition of their untranslated regions. By sequence comparison, we have mapped the novel gene, named S100A15, to the S100 gene cluster within the epidermal differentiation complex ( chromosome 1q21). Analysis of the deduced amino acid sequence revealed a protein of 101 amino acids containing two potential EF-hand motifs with high homology to the S100A7. Northern blot hybridization and semiquantitative RT-PCR analysis confirmed the S100A15 overexpression in psoriasis, showing different levels of expression of the S100A15 mRNA isoforms. In situ hybridization of the S100A15 revealed a markedly increased staining of basal and suprabasal epidermal layers of psoriatic skin compared with healthy tissue. Our data suggest an involvement of the novel S100A15 in epidermal differentiation and inflammation and might therefore be important for the pathogenesis of psoriasis and other diseases.