• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Regulation of nucleocytoplasmic localization of the atDjC6 chaperone protein
【24h】

Regulation of nucleocytoplasmic localization of the atDjC6 chaperone protein

机译：atDjC6伴侣蛋白的核质定位调节

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The sequence of the atDjC6 chaperone protein includes three potential nuclear localization signal (NLS) sequences (A-C) and three potential nuclear export signal (NES) sequences (X-Z). The subcellular localization of atDjC6 was studied by scanning laser confocal microscopy of chimera with the green-fluorescent protein (GFP) transiently expressed in tobacco BY-2 cells. The localization of the atDjC6::GFP chimera was coincident with that of the nuclear stain propidium iodide. Site-directed mutagenesis was used to verify the predicted NLS sequences. Each was individually fused to GFP and tested for protein localization. The individual NLS sequences were sufficient to direct partial nuclear localization of GFP, although the targeting information within NLS-B is apparently conformation sensitive. Site-directed mutagenesis of the NES sequences increased the amount of each chimera that was nuclearly localized, indicating a decrease in nuclear export. When any pair of NLS sequences were appended to GFP, the chimera were entirely nuclearly localized. Quantitative two-hybrid analysis was used to verify that the decoding of NLS sequence information involves interaction with the NLS-receptor protein importin-alpha. Each of the NLS sequences is flanked by a site of potential Ser phosphorylation, and recombinant atDjC6 could be phosphorylated in vitro. Mutagenesis of Ser residues to the P-Ser mimic Asp interfered with nuclear targeting, apparently by preventing recognition or binding by importin-alpha. Our results are consistent with a regulated nucleocytoplasmic localization of the atDjC6 chaperone protein.
机译：atDjC6伴侣蛋白的序列包括三个潜在的核定位信号（NLS）序列（A-C）和三个潜在的核输出信号（NES）序列（X-Z）。通过扫描嵌合体的激光共聚焦显微镜对atDjC6的亚细胞定位进行了研究，其在烟草BY-2细胞中瞬时表达了绿色荧光蛋白（GFP）。 atDjC6 :: GFP嵌合体的定位与核染色剂碘化丙啶的定位一致。定点诱变用于验证预测的NLS序列。每个都单独融合到GFP，并测试蛋白质的定位。尽管NLS-B中的靶向信息显然对构象敏感，但单个NLS序列足以指导GFP的部分核定位。 NES序列的定点诱变增加了每个嵌合核定位的数量，表明核输出减少。当将任意一对NLS序列附加到GFP时，嵌合体完全在核中定位。定量两杂交分析用于验证NLS序列信息的解码涉及与NLS受体蛋白importin-alpha的相互作用。每个NLS序列的侧面都带有潜在的Ser磷酸化位点，重组atDjC6可以在体外被磷酸化。 Ser残基突变为P-Ser模拟物Asp显然干扰了importin-alpha的识别或结合，从而干扰了核靶向。我们的结果与atDjC6伴侣蛋白的核仁定位调控相一致。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号