TdT interacting factor 1 enhances TdT ubiquitylation through recruitment of BPOZ-2 into nucleus from cytoplasm
TdT interacting factor 1 enhances TdT ubiquitylation through recruitment of BPOZ-2 into nucleus from cytoplasm
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DOI：
10.1111/j.1365-2443.2009.01358.x
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发表时间：
2009-12-01
期刊：
影响因子：
2.1
通讯作者：
Koiwai, Osamu
中科院分区：
文献类型：
--
作者：
Hayano, Takahide;Koiwai, Kotaro;Koiwai, Osamu
我们使用酵母双杂交系统分离了编码含有2型基因（BPOZ-2）的人cDNA克隆，该基因具有与TdT相互作用因子1（TdIF 1）结合的产物。BPOZ-2是E3连接酶CUL 3的接头，参与发育过程。BPOZ-2和TdIF 1之间的结合，证实了GST下拉和免疫沉淀试验，使用特异性抗体对BPOZ-2和TdIF 1在体外和体内。虽然当BPOZ-2在COS 7细胞中单独表达时，观察到BPOZ-2主要在细胞质中，但将pEGFP-BPOZ-2和pDsRed-TdIF 1共转染入COS 7细胞中导致EGFP-BPOZ-2和DsRed-TdIF 1共定位于细胞核中。TdIF 1可以通过直接结合BPOZ-2将BPOZ-2从细胞质募集到细胞核中。当TdIF 1与BPOZ-2在293 T细胞中一起表达时，BPOZ-2增强TdT泛素化，强烈表明BPOZ-2从细胞质募集到细胞核中对于细胞核内的TdT泛素化是重要的。
We isolated human cDNA clone encoding Bood POZ containing gene type 2 (BPOZ-2) as a gene with a product that binds to TdT interacting factor 1 (TdIF1) using a yeast two-hybrid system. BPOZ-2 is an adaptor for E3 ligase CUL3 and participates in developmental processes. The binding between BPOZ-2 and TdIF1 was confirmed by GST pull-down and immunoprecipitation assays using specific antibodies against BPOZ-2 and TdIF1 in vitro and in vivo. Although when BPOZ-2 solely was expressed in COS7 cells, BPOZ-2 was observed mainly within the cytoplasm, co-transfection of pEGFP-BPOZ-2 and pDsRed-TdIF1 into COS7 cells resulted in co-localization of EGFP-BPOZ-2 and DsRed-TdIF1 within the nucleus. TdIF1 may recruit BPOZ-2 into the nucleus from the cytoplasm by directly binding to BPOZ-2. BPOZ-2 enhanced TdT ubiquitylation when TdIF1 was expressed together with BPOZ-2 in 293T cells, strongly suggesting that the recruitment of BPOZ-2 into the nucleus from the cytoplasm is significant for the TdT ubiquitylation within the nucleus.