Zebrafish Dnd protein binds to 3'UTR of geminin mRNA and regulates its expression
Zebrafish Dnd protein binds to 3'UTR of geminin mRNA and regulates its expression
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DOI：
10.5483/bmbrep.2010.43.6.438
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发表时间：
2010-06-30
期刊：
影响因子：
3.8
通讯作者：
Ma, Yongxin
中科院分区：
文献类型：
--
作者：
Chen, Shu;Zeng, Mei;Ma, Yongxin
Dnd（dead end）基因编码一种RNA结合蛋白，并在原始生殖细胞（PGCs）中特异性表达，作为整个胚胎发生过程中生殖细胞浆的脊椎动物特异性组分。利用特异性核酸与蛋白质结合技术（SNAAP），从斑马鱼8-细胞胚胎中鉴定出13个ZDnd蛋白的潜在靶mRNA，其中8个靶mRNA经RT-PCR分析得到确认。在靶mRNA中，本研究的重点是对DNA复制抑制剂geminin的调控。电泳迁移率变动分析（EMSA）结果表明，ZDND蛋白与gemininmRNA的3 'UTR中864 ~ 931位67个核苷酸的区域结合，该区域含有60.29%的尿苷。来自HEK 293细胞中的双荧光素酶测定的结果表明，ZDND增加了双生蛋白的翻译。总之，ZDnd靶mRNA的鉴定将有助于进一步探讨Dnd在斑马鱼种系发育以及癌细胞中的生物学功能。[BMB报告2010; 43（6）：438-444]
Dnd (dead end) gene encodes an RNA binding protein and is specifically expressed in primordial germ cells (PGCs) as a vertebrate-specific component of the germ plasma throughout embryogenesis. By utilizing a technique of specific nucleic acids associated with proteins (SNAAP), 13 potential target mRNAs of zebrafish Dnd (ZDnd) protein were identified from 8-cell embryo, and 8 target mRNAs have been confirmed using an RT-PCR analysis. Of the target mRNAs, the present study is focused on the regulation of geminin, which is an inhibitor of DNA replication. Using electrophoretic mobility shift assay (EMSA), we demonstrated that ZDND protein bound the 67-nucleotide region from 864 to 931 in the 3'UTR of geminin mRNA, a sequence containing 60.29% of uridine. Results from a dual-luciferase assay in HEK293 cells showed that ZDND increases the translation of geminin. Taken together, the identification of target mRNA for ZDnd will be helpful to further explore the biological function of Dnd in zebrafish germ-line development as well as in cancer cells. [BMB reports 2010; 43(6): 438-444]