Transcriptional Regulation of Vih by Oct4 and Sox9 in Scylla paramamosain.
Transcriptional Regulation of Vih by Oct4 and Sox9 in Scylla paramamosain.
复制标题
Oct4 和 Sox9 对拟青青蟹 Vih 的转录调控
DOI：
10.3389/fendo.2020.00650
复制
发表时间：
2020
影响因子：
5.2
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Liao J;Zhang Z;Jia X;Zou Z;Liang K;Wang Y
青蟹(Scylla Paramosain)是中国最具经济价值的海蟹之一。然而，对生殖调节机制的研究还不够充分。卵黄发生抑制激素(VIH)是甲壳动物高血糖激素(CHH)家族的一员，在甲壳动物性腺发育和成熟过程中起着重要的调节作用，而目前对VIH转录调控的研究相对较少。我们以前对泥蟹VIH(SpVih)转录调控的研究已经证明，Oct4/Sox9转录因子的结合部位可能是正向调控SpVih表达的关键区域。在本研究中，凝胶迁移率改变分析(EMSA)实验证实，从眼柄提取的核蛋白可以与SpVih启动子的关键区域结合，并且这些特异结合依赖于Oct4/Sox9结合位点的存在。用Oct4和Sox9抗体在EMSA超移位实验的超移组中检测到两条特异性结合复合带，进一步证实了这两种转录因子对SpVih关键调控区的特异性识别。体外构建Oct4、Sox9基因过表达载体和SpVih核心启动子片段载体，共转染HEK293T细胞。结果表明，SpVih活性随转录因子浓度的增加而增加。在体内，分别向眼柄注射Oct4和Sox9 dsRNA后，SpVih在Oct4或Sox9干预后的表达水平显著降低，而SPVTG在卵巢和肝胰腺中的表达水平显著升高。体外和体内实验均表明，Oct4和Sox9对SpVih有正向调节作用。用纯化的Oct4和Sox9蛋白进行GST下拉实验，结果表明它们之间存在相互作用。推测它们通过相互作用调节SpVih的表达。
Mud crab (Scylla paramamosain) is one of the most economically-important marine crabs in China. However, research on mechanisms of reproductive regulation is not sufficient. Vitellogenesis-inhibiting hormone (VIH) is a member of the crustacean hyperglycemia hormones (CHH) family, which plays an essential role in the regulation of gonadal development and maturation in crustaceans, and current studies on the regulation of Vih transcription in crabs are relatively rare. Our previous studies on the transcriptional regulation of mud crab Vih (SpVih) have proved that the binding site of Oct4/Sox9 transcription factor may be the key region for positively regulating the expression of SpVih. In this study, the electrophoretic mobility shift assay (EMSA) experiment confirmed that the nuclear protein extracted from the eyestalk could bind to the key region of SpVih promoter, and these specific bindings were dependent on the presence of Oct4/Sox9 binding sites. Two specific binding complex bands were detected in the supershift group of EMSA supershift experiments by Oct4 and Sox9 antibodies, further confirming the specific recognition of these two transcription factors on the key regulatory region of SpVih. In vitro, Oct4 and Sox9 gene overexpression vectors and SpVih core promoter fragment vector were constructed and co-transfected into HEK293T cells. As a result, SpVih activity increased with the concentration of transcription factors. In vivo, when Oct4 and Sox9 dsRNA were injected into the eyestalks of mud crab, respectively, the expression level of SpVih decreased significantly after interference with Oct4 or Sox9, and the expression level of SpVtg in the ovary and hepatopancreatic increased. Both in vitro and in vivo experiments showed that Oct4 and Sox9 had a positive regulatory effect on SpVih. The GST pull-down experiment was carried out by purified Oct4 and Sox9 proteins, and the results showed that there was an interaction between them. It was speculated that they regulated the expression of SpVih through the interaction.
登录
查看更多内容
影响因子：
37.3
作者：
Jen J;Tang YA;Lu YH;Lin CC;Lai WW;Wang YC
通讯作者：
Wang YC
影响因子：
2.3
作者：
Nanda, S.;DeFalco, T. J.;Russell, S.
通讯作者：
Russell, S.
TGF-β1-SOX9轴诱导的COL10A1通过上皮间质转化促进胃癌侵袭和转移
影响因子：
9
作者：
Li T;Huang H;Shi G;Zhao L;Li T;Zhang Z;Liu R;Hu Y;Liu H;Yu J;Li G
通讯作者：
Li G
影响因子：
4.5
作者：
Das, Rekha;Krishna, Gopal;Chaudhari, Aparna
通讯作者：
Chaudhari, Aparna
影响因子：
2.7
作者：
Barske, Lindsey A.;Capel, Blanche
通讯作者：
Capel, Blanche