STAT4DNA结合域中395~416位氨基酸残基序列参与.pdfVIP

生理学报 Acta Physiologica Sinica, August 25, 2012, 64(4): 372–378 372 Received 2012-04-26 Accepted 2012-07-02 This work was supported by the National Natural Scientific Foundation of China (No. and National Undergraduate Innovation Training Project of Ministry of Education, China (No. AE11528). *Corresponding author. Tel: +86-731 E-mail: liming@ STAT4的DNA结合域中395~416位氨基酸残基序列参与IL-12 介导的STAT4入核 黄玉梅1，文亚平1，李轩岸2，袁 媛1，罗奇志1，黎 明1,* 中南大学湘雅医学院1 基础医学院免疫学教研室； 2 临床医学五年制07级教改班，长沙 410078 摘 要：本研究旨在探讨信号转导和转录活化蛋白4 (signal transducer and activator of transcription 4, STAT4)受白介素-12 (in- terleukin-12, IL-12)刺激后移位入细胞核的机制。对STATs家族成员进行同源性比对分析结果显示，STAT4的DNA结合域的 395~416位氨基酸残基序列可能具有二聚体特异性核定位信号(dimer-specific nuclear localization signal, dsNLS)功能。有鉴于 此，本研究以pEGFP-C1为表达载体，分别构建了pEGFP-STAT4质粒、缺失395~416位氨基酸残基序列的缺失型STAT4质粒 (pEGFP-STAT4-Del)、将SV40大T抗原上经典的NLS核酸序列插入表达载体的阳性对照质粒(pEGFP-NLS)和将缺失型STAT4 插入pEGFP-NLS的pEGFP-NLS-STAT4-Del质粒。将这些质粒瞬时转染宫颈癌腺癌细胞系Caski细胞，经过IL-12刺激，发现 野生型STAT4移位入细胞核，而缺失型STAT4分布在细胞浆中；进一步用leptomycin B处理，IL-12再刺激后野生型STAT4被 滞留于细胞核中，而缺失型STAT4仍然分布在胞浆中；将NLS插入缺失型STAT4，能恢复缺失型STAT4的核移位。以上结 果说明野生型STAT4在IL-12刺激下能移位入细胞核，其入核机制是在其DNA结合域的395~416位氨基酸残基序列具有 dsNLS功能，能介导活化的STAT4移位入细胞核。 关键词：核移位；核定位信号；信号转导和转录活化蛋白4 中图分类号：R3 Amino acids 395–416 in DNA binding domain of STAT4 is involved in IL-12-induced nuclear import of STAT4 HUANG Yu-Mei1, WEN Ya-Ping1, LI Xuan-An2, YUAN Yuan1, LUO Qi-Zhi1, LI Ming1,* 1Department of Immunology, Basic Medical College; 2Teaching Innovation Class of Clinical Medicine Five-year Program 07 Grade, Xiangya Medical College, Central South University, Changsha 410078, China Abstract: The purpose of the present study is to explore the mechanism of IL-12-induced nuclear import of signal transducer and activator of transcription 4 (STAT4). Assayed by analyses of homology alignment of STATs, amino acids 395–416 in DNA binding domain was found to be a potential dimer-specific nuclear localization signal (dsNLS) of STAT4. Therefore, several plasmids were construct