T-box transcription factor eomesodermin/Tbr2 in Atlantic cod (Gadus morhua L.): Molecular characterization, promoter structure and function analysis
T-box transcription factor eomesodermin/Tbr2 in Atlantic cod (Gadus morhua L.): Molecular characterization, promoter structure and function analysis
复制标题
大西洋鳕鱼 (Gadus morhua L.) 中的 T 盒转录因子 eomesodermin/Tbr2：分子表征、启动子结构和功能分析
DOI：
10.1016/j.fsi.2019.07.021
复制
发表时间：
2019
影响因子：
4.7
通讯作者：
Dalmo Roy Ambli
中科院分区：
文献类型：
--
作者：
Chi Heng;Sormo Kristian Gillebo;Diao Jing;Dalmo Roy Ambli
Eomesodermin(Eomesodermin，Eomesodermin)是T-box转录因子家族的一员，在动物多种发育过程和免疫反应的调节中发挥着重要作用。在这里，我们报道了大西洋鳕鱼全长cDNA(GmEome)的克隆和鉴定，该基因具有与哺乳动物类似的Tbox_3结构域。在活体感染鳗弧菌后，头肾和脾中有表达上调，PMA和Poly I：C刺激后，脾中有白细胞表达上调。此外，我们在转录起始点(TSS)上游确定了一个694bp的序列，包含许多与已知转录因子结合位点匹配的序列基序。通过在CHSE-214 细胞中导入不同的5‘-缺失结构来检测推测的调控基因的活性。结果表明，GmEome的基本启动子和正转录调节活性受−基因上游694~−376bp的序列控制。此外，我们还发现在生物信息学预测的鳕鱼干扰素γ基因的5‘侧翼区存在一些eome结合位点。然而，将eomesodermin高表达载体与INFγ报告载体共转染CHSE-214 细胞，结果表明大西洋鳕鱼eomesodermin在INFγ启动子的激活中起到了次要作用。
Eomesodermin (Eomes) is a member of T-box transcription factor family and plays an important role in the regulation of a wide variety of developmental processes and immune response in animals. Here we report cloning and characterization of the full-length cDNA of Atlantic cod Eomes (GmEomes), which possesses a TBOX_3 domain similar to its counterpart in mammals. The regulated expression was observed in head kidney and spleen in response to liveVibrio anguillaruminfectionin vivo, and spleen leukocytesin vitroafter PMA and poly I:C stimulation. Furthermore, we determined a 694 bp sequence, upstream of the transcriptional start site (TSS), to contain a number of sequence motifs that matched known transcription factor-binding sites. Activities of the presumptive regulatory gene were assessed by transfecting different 5′-deletion constructs in CHSE-214 cells. The results showed that the basal promoters and positive transcriptional regulator activities of GmEomes were dependent by sequences located from −694 to −376 bp upstream of TSS. Furthermore, we found that some Eomes binding sites were present in the 5′-flanking regions of the cod IFNγ gene predicted by bioinformatics. However, Co-transfection of eomesodermin overexpression plasmids with INFγ reporter vector into CHSE-214 cells determined that Atlantic cod eomesodermin played a minor role in activation of the INFγ promoter.