Cytoplasmic relocation of Daxx induced by Ro52 and FLASH.
Cytoplasmic relocation of Daxx induced by Ro52 and FLASH.
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DOI：
10.1007/s00418-010-0734-6
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发表时间：
2010-09
影响因子：
2.3
通讯作者：
Kamitani, Tetsu
中科院分区：
文献类型：
--
作者：
Tanaka, Makoto;Kamitani, Tetsu
已知环指蛋白Ro 52/TRIM 21是一种自身抗原，可被抗Ro/SSA抗体识别，这种抗体常见于干燥综合征和系统性红斑狼疮患者中。我们最近发现Ro 52是一种E3泛素连接酶，定位于沿着微管网络高度运动的细胞质体。为了扩大我们对Ro 52的了解，我们寻找了与Ro 52合作的合作伙伴。以Ro 52为诱饵，对人脑cDNA文库进行酵母双杂交筛选。该筛选确定了几个编码Ro 52相互作用蛋白的基因，包括乳腺癌相关蛋白Daxx和FLASH。进一步的酵母双杂交试验显示，Daxx结合Ro 52的B30.2结构域，FLASH通过其死亡效应结构域募集结构域结合Ro 52的卷曲螺旋结构域。这些结果表明Ro 52、Daxx和FLASH形成异聚蛋白复合物。事实上，这得到了免疫沉淀实验结果的支持，我们发现在过表达的FLASH存在下，Daxx与Ro 52共免疫沉淀。重要的是，我们的荧光显微镜显示，虽然Daxx主要位于细胞核中，但Ro 52和FLASH的过表达导致Daxx重新定位到细胞质中。因此，Ro 52似乎与FLASH合作以诱导Daxx在细胞中的细胞质定位。
The RING-finger protein Ro52/TRIM21 is known to be an autoantigen and is recognized by anti-Ro/SSA antibodies, which are commonly found in patients with Sjögren's syndrome and systemic lupus erythematosus. We recently showed that Ro52 is an E3 ubiquitin ligase and localizes to cytoplasmic bodies that are highly motile along the microtubule network. To expand our knowledge of Ro52, we searched partners co-operating with Ro52. We performed a yeast two-hybrid screening of a human brain cDNA library with Ro52 as bait. This screening identified several genes encoding Ro52-interacting proteins, including the apoptosis-related proteins, Daxx and FLASH. Further yeast two-hybrid assays revealed that Daxx binds to the B30.2 domain of Ro52 and that FLASH binds to coiled-coil domains of Ro52 through its death-effector domain-recruiting domain. These results suggest that Ro52, Daxx, and FLASH form heteromeric protein complexes. Indeed, this was supported by results of immunoprecipitation experiments in which we found that Daxx is co-immunoprecipitated with Ro52 in the presence of overexpressed FLASH. Importantly, our fluorescence microscopy revealed that, although Daxx is predominantly located in the nucleus, overexpression of both Ro52 and FLASH leads to relocation of Daxx into the cytoplasm. Thus, Ro52 seems to co-operate with FLASH to induce cytoplasmic localization of Daxx in cells.
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影响因子：
11.4
作者：
Milovic-Holm, Kristijana;Krieghoff, Eva;Hofmann, Thomas G.
通讯作者：
Hofmann, Thomas G.
DOI：
10.1016/j.bbrc.2006.01.144
发表时间：
2006-03-31
影响因子：
3.1
作者：
Wada, K;Kamitani, T
通讯作者：
Kamitani, T
DOI：
10.1016/s1357-2725(01)00055-3
发表时间：
2001-09-01
影响因子：
4
作者：
Di Donato, F;Chan, EKL;Buyon, JP
通讯作者：
Buyon, JP
DOI：
10.1016/j.bbrc.2005.11.076
发表时间：
2006-01-20
影响因子：
3.1
作者：
Wada, K;Tanji, K;Kamitani, T
通讯作者：
Kamitani, T
影响因子：
5.4
作者：
Yamauchi, Keiko;Wada, Keiji;Kamitani, Tetsu
通讯作者：
Kamitani, Tetsu