The synovial sarcoma associated protein SYT interacts with the acute leukemia associated protein AF10
The synovial sarcoma associated protein SYT interacts with the acute leukemia associated protein AF10
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DOI：
10.1038/sj.onc.1204419
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发表时间：
2001-05-31
期刊：
影响因子：
8
通讯作者：
van Kessel, AG
中科院分区：
文献类型：
--
作者：
de Bruijn, DRH;dos Santos, NR;van Kessel, AG
由于滑膜肉瘤相关的t（X;18）易位，人类18号染色体上的SYT基因与X染色体上的SSX1或SSX2基因融合。虽然初步证据表明，这些基因编码的（融合）蛋白可能在转录调控中发挥作用，但对其确切功能知之甚少。我们以SYT为诱饵，通过酵母双杂交筛选人类cDNA文库分离相互作用蛋白。在分离的阳性克隆中，有两个被发现与急性白血病t（10;11）相关的AF10基因相对应，AF10基因是MLL的融合伙伴，通过细胞系提取物中内源性和外源性表位标记的SYT和AF10蛋白的共免疫沉淀以及表位标记的SYT和AF10蛋白在转染细胞中的共定位，证实了这些结果。随后的序列突变分析显示，n端SYT片段与c端AF10片段具有高度特异性的相互作用，SYT蛋白的n端相互作用域也存在于几个SYT同源物和同源物中，AF10的c端相互作用域位于已知功能域之外。基于这些结果，提出了SYT和AF10蛋白作为双部转录因子协同作用的模型。该模型对人类滑膜肉瘤和急性白血病发展的分子机制具有启示意义。
As a result of the synovial sarcoma associated t(X;18) translocation, the human SYT gene on chromosome 18 is fused to either the SSX1 or the SSX2 gene on the X chromosome. Although preliminary evidence indicates that the (fusion) proteins encoded by these genes may play a role in transcriptional regulation, little is known about their exact function. We set out to isolate interacting proteins through yeast two hybrid screening of a human cDNA library using SYT as a bait. Of the positive clones isolated, two were found to correspond to the acute leukemia t(10;11) associated AF10 gene, a fusion partner of MLL, Confirmation of these results was obtained via co-immunoprecipitation of endogenous and exogenous, epitope-tagged, SYT and AF10 proteins from cell line extracts and colocalization of epitope-tagged SYT and AF10 proteins in transfected cells. Subsequent sequential mutation analysis revealed a highly specific interaction of N-terminal SYT fragments with C-terminal AF10 fragments, The N-terminal interaction domain of the SYT protein was also found to be present in several SYT orthologs and homologs, The C-terminal interaction domain of AF10 is located outside known functional domains. Based on these results, a model is proposed in which the SYT and AF10 proteins act in concert as bipartite transcription factors. This model has implications for the molecular mechanisms underlying the development of both human synovial sarcomas and acute leukemias.