Radoja Nada, Stojadinovic Olivera, Waseem Ahmad, Tomic-Canic Marjana, Milisavljevic Vladana, Teebor Susan, Blumenberg Miroslav
Department of Dermatology, New York University School of Medicine, New York, New York 10016, USA.
Mol Cell Biol. 2004 Apr;24(8):3168-79. doi: 10.1128/MCB.24.8.3168-3179.2004.
Basal layers of stratified epithelia express keratins K5, K14, and K15, which assemble into intermediate filament networks. Mutations in K5 or K14 genes cause epidermolysis bullosa simplex (EBS), a disorder with blistering in the basal layer due to cell fragility. Nonkeratinizing stratified epithelia, e.g., in the esophagus, produce more keratin K15 than epidermis, which alleviates the esophageal symptoms in patients with K14 mutations. Hypothesizing that increasing the cellular content of K15 could compensate for the mutant K14 and thus ease skin blistering in K14 EBS patients, we cloned the promoter of the K15 gene and examined its transcriptional regulation. Using cotransfection, gel mobility shifts, and DNase I footprinting, we have identified the regulators of K15 promoter activity and their binding sites. We focused on those that can be manipulated with extracellular agents, transcription factors C/EBP, AP-1, and NF-kappaB, nuclear receptors for thyroid hormone, retinoic acid, and glucocorticoids, and the cytokine gamma interferon (IFN-gamma). We found that C/EBP-beta and AP-1 induced, while retinoic acid, glucocorticoid receptors, and NF-kappaB suppressed, the K15 promoter, along with other keratin gene promoters. However, the thyroid hormone and IFN-gamma uniquely and potently activated the K15 promoter. Using these agents, we could boost the amounts of K15 in human epidermis. Our findings suggest that treatments based on thyroid hormone and IFN-gamma could become effective agents in therapy for patients with EBS.
复层上皮的基底层表达角蛋白K5、K14和K15,它们组装成中间丝网络。K5或K14基因的突变会导致单纯性大疱性表皮松解症(EBS),这是一种由于细胞脆弱而在基底层出现水疱的疾病。非角化复层上皮,如食管中的上皮,产生的角蛋白K15比表皮更多,这减轻了K14突变患者的食管症状。我们推测增加K15的细胞含量可以补偿突变的K14,从而缓解K14型EBS患者的皮肤水疱症状,于是我们克隆了K15基因的启动子并研究其转录调控。通过共转染、凝胶迁移率变动分析和DNase I足迹分析,我们确定了K15启动子活性的调节因子及其结合位点。我们重点研究了那些可以用细胞外因子调控的调节因子,即转录因子C/EBP、AP-1和NF-κB、甲状腺激素、视黄酸和糖皮质激素的核受体以及细胞因子γ干扰素(IFN-γ)。我们发现C/EBP-β和AP-1诱导K15启动子,而视黄酸、糖皮质激素受体和NF-κB抑制K15启动子以及其他角蛋白基因启动子。然而,甲状腺激素和IFN-γ独特且强力地激活K15启动子。使用这些因子,我们可以提高人表皮中K15的含量。我们的研究结果表明,基于甲状腺激素和IFN-γ的治疗方法可能成为治疗EBS患者的有效药物。