Dynein Light Chain 1 (DYNLT1) Interacts with Normal and Oncogenic Nucleoporins.
Dynein Light Chain 1 (DYNLT1) Interacts with Normal and Oncogenic Nucleoporins.
复制标题
DOI：
10.1371/journal.pone.0067032
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Yaseen NR
中科院分区：
文献类型：
--
作者：
Sarma NJ;Yaseen NR
嵌合癌蛋白NUP 98-HOXA 9由t（7;11）（p15;p15）染色体易位产生，并且与急性髓性白血病相关。它通过尚未完全理解的机制引起异常基因调控和白血病转化。NUP 98-HOXA 9由核孔蛋白NUP 98的N-末端部分组成，其含有许多与HOXA 9的DNA结合同源结构域融合的FG重复序列。我们使用Cytotrap酵母双杂交测定来鉴定与NUP 98-HOXA 9相互作用的蛋白质。我们确定了动力蛋白轻链1（DYNLT 1），一个完整的14 KDa蛋白亚基的大微管为基础的细胞质动力蛋白复合物，作为NUP 98-HOXA 9的相互作用伴侣。通过体外下拉和免疫共沉淀测定证实结合，并且NUP 98-HOXA 9的FG重复区显示为相互作用所必需的。RNAi介导的DYNLT 1敲低导致NUP 98-HOXA 9激活转录的能力降低，并且还抑制NUP 98-HOXA 9诱导原代人造血CD 34+细胞增殖的能力。DYNLT 1还显示出与野生型NUP 98和其他含有FG重复的核孔蛋白的强相互作用。免疫荧光分析表明，DYNLT 1主要定位于核周边，在那里它与核孔复合物共定位，并定位于细胞质。缺失研究表明，核孔蛋白与DYNLT 1的相互作用主要依赖于DYNLT 1的C-末端的一半。这些数据首次表明DYNLT 1与核孔蛋白相互作用，并在基因表达失调和通过致白血病核孔蛋白融合NUP 98-HOXA 9诱导造血细胞增殖中发挥作用。
The chimeric oncoprotein NUP98-HOXA9 results from the t(7;11)(p15;p15) chromosomal translocation and is associated with acute myeloid leukemia. It causes aberrant gene regulation and leukemic transformation through mechanisms that are not fully understood. NUP98-HOXA9 consists of an N-terminal portion of the nucleoporin NUP98 that contains many FG repeats fused to the DNA-binding homeodomain of HOXA9. We used a Cytotrap yeast two-hybrid assay to identify proteins that interact with NUP98-HOXA9. We identified Dynein Light Chain 1 (DYNLT1), an integral 14 KDa protein subunit of the large microtubule-based cytoplasmic dynein complex, as an interaction partner of NUP98-HOXA9. Binding was confirmed by in vitro pull down and co-immunoprecipitation assays and the FG repeat region of NUP98-HOXA9 was shown to be essential for the interaction. RNAi-mediated knockdown of DYNLT1 resulted in reduction of the ability of NUP98-HOXA9 to activate transcription and also inhibited the ability of NUP98-HOXA9 to induce proliferation of primary human hematopoietic CD34+ cells. DYNLT1 also showed a strong interaction with wild-type NUP98 and other nucleoporins containing FG repeats. Immunofluorescence analysis showed that DYNLT1 localizes primarily to the nuclear periphery, where it co-localizes with the nuclear pore complex, and to the cytoplasm. Deletion studies showed that the interactions of the nucleoporins with DYNLT1 are dependent predominantly on the C-terminal half of the DYNLT1. These data show for the first time that DYNLT1 interacts with nucleoporins and plays a role in the dysregulation of gene expression and induction of hematopoietic cell proliferation by the leukemogenic nucleoporin fusion, NUP98-HOXA9.
登录
查看更多内容
影响因子：
5.3
作者：
Galigniana, Mario D.;Erlejman, Alejandra G.;Piwien-Pilipuk, Graciela
通讯作者：
Piwien-Pilipuk, Graciela
影响因子：
25
作者：
Gauthier-Fisher, Andree;Lin, Dan C.;Miller, Freda D.
通讯作者：
Miller, Freda D.
影响因子：
3.5
作者：
Jiang, JM;Yu, L;Zhao, SY
通讯作者：
Zhao, SY
影响因子：
2.9
作者：
Ma, Xianyue;Sayeski, Peter P.
通讯作者：
Sayeski, Peter P.
影响因子：
11.8
作者：
Chuang, JZ;Yeh, TY;Sung, CH
通讯作者：
Sung, CH