Functional and structural basis of the nuclear localization signal in the ZIC3 zinc finger domain.
Functional and structural basis of the nuclear localization signal in the ZIC3 zinc finger domain.
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DOI：
10.1093/hmg/ddn239
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发表时间：
2008-11-15
影响因子：
3.5
通讯作者：
Aruga, Jun
中科院分区：
文献类型：
--
作者：
Hatayama, Minoru;Tomizawa, Tadashi;Sakai-Kato, Kumiko;Bouvagnet, Patrice;Kose, Shingo;Imamoto, Naoko;Yokoyama, Shigeyuki;Utsunomiya-Tate, Naoko;Mikoshiba, Katsuhiko;Kigawa, Takanori;Aruga, Jun
ZIC 3的破坏会导致异位，这是一种先天性左右轴异常。ZIC 3编码一个具有锌指结构域的核蛋白，该锌指结构域包含5个串联的C2 H2 ZF基序。错义突变在第一ZF基序（ZF 1）导致缺陷的核定位，这可能是基础的异位症的发病机制。本研究揭示了ZIC 3核定位信号（NLS）的结构和功能基础，并探讨了其与ZF 1突变缺陷的关系。ZIC 3 NLS位于ZF 2和ZF 3区域，而不是ZF 1。散布在这些区域的几个碱性残基负责核定位，但R320，K337和R350是特别重要的。核磁共振结构分析表明，ZF 1 -4具有与GLI ZF相似的结构，NLS的碱性侧链聚集在蛋白质表面的两个区域，类似于经典的二分NLS。在用于ZF 1突变的残基中，C253和H286被定位用于金属螯合，而W255被定位在由ZF 1和ZF 2形成的疏水核心中。色氨酸255是一个高度保守的指间连接物，并形成与GLI、Glis和一些真菌ZF蛋白共享的结构基序（串联CXW-C-H-H）的一部分。此外，我们发现敲低Karyopherin α1/α6会损害ZIC 3的核定位，NLS与核输入衔接蛋白之间的物理相互作用会受到NLS突变的干扰，但不会受到W255 G的干扰。这些结果表明，ZIC 3是进口到细胞核的Karyopherin（Importin）系统和受损的核定位的ZF 1突变是由于对NLS的直接影响。
Disruptions in ZIC3 cause heterotaxy, a congenital anomaly of the left–right axis. ZIC3 encodes a nuclear protein with a zinc finger (ZF) domain that contains five tandem C2H2 ZF motifs. Missense mutations in the first ZF motif (ZF1) result in defective nuclear localization, which may underlie the pathogenesis of heterotaxy. Here we revealed the structural and functional basis of the nuclear localization signal (NLS) of ZIC3 and investigated its relationship to the defect caused by ZF1 mutation. The ZIC3 NLS was located in the ZF2 and ZF3 regions, rather than ZF1. Several basic residues interspersed throughout these regions were responsible for the nuclear localization, but R320, K337 and R350 were particularly important. NMR structure analysis revealed that ZF1–4 had a similar structure to GLI ZF, and the basic side chains of the NLS clustered together in two regions on the protein surface, similar to classical bipartite NLSs. Among the residues for the ZF1 mutations, C253 and H286 were positioned for the metal chelation, whereas W255 was positioned in the hydrophobic core formed by ZF1 and ZF2. Tryptophan 255 was a highly conserved inter-finger connector and formed part of a structural motif (tandem CXW-C-H-H) that is shared with GLI, Glis and some fungal ZF proteins. Furthermore, we found that knockdown of Karyopherin α1/α6 impaired ZIC3 nuclear localization, and physical interactions between the NLS and the nuclear import adapter proteins were disturbed by mutations in the NLS but not by W255G. These results indicate that ZIC3 is imported into the cell nucleus by the Karyopherin (Importin) system and that the impaired nuclear localization by the ZF1 mutation is not due to a direct influence on the NLS.
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影响因子：
56.9
作者：
Carninci, P;Kasukawa, T;Hayashizaki, Y
通讯作者：
Hayashizaki, Y
影响因子：
14.9
作者：
Beak, Ju Youn;Kang, Hong Soon;Kim, Yong-Sik;Jetten, Anton M.
通讯作者：
Jetten, Anton M.
影响因子：
5.6
作者：
Fontes, MRM;Teh, T;Kobe, B
通讯作者：
Kobe, B
影响因子：
4.8
作者：
Fontes, MRM;Teh, T;Kobe, B
通讯作者：
Kobe, B
影响因子：
4
作者：
Cutress, Mark L.;Whitaker, Hayley C.;Neal, David E.
通讯作者：
Neal, David E.