血蓝蛋白,一种含铜的呼吸蛋白,大量存在于节肢动物和软体动物的血淋巴中,并具有多种免疫功能。然而,血蓝蛋白基因转录的调控机制尚不清楚。我们以前的工作表明,敲低转录因子CSL,Notch信号通路的一个组成部分,下调南美白对虾血蓝蛋白小亚基基因(PvHMCs)的表达,表明CSL参与调节PvHMCs转录。在这项研究中,我们确定了一个CSL结合基序(“GAATCCCAGA”,PvHMCs(命名为HsP3)的核心启动子中的+1675/+1684bp)。双荧光素酶报告基因测定和电泳迁移率变化测定(EMSA)表明凡纳滨对虾的CSL同源物(PvCSL)可以直接结合并激活HsP3启动子。此外,PvCSL的体内沉默显著减弱了PvHMCs的mRNA和蛋白表达。最后,对副溶血性弧菌的反应,链球菌和白斑综合征病毒(WSSV)攻击,PvCSL和PvHMCs的转录本呈正相关,提示PvCSL还可以在病原体刺激下调节PvHMCs的表达。一起来看,我们目前的发现是第一个证明PvCSL是PvHMCs转录控制的关键因素。
Hemocyanin, a copper-containing respiratory protein, is abundantly present in hemolymph of arthropods and mollusks and performs a variety of immunological functions. However, the regulatory mechanisms of hemocyanin gene transcription remain largely unclear. Our previous work showed that knockdown of the transcription factor
CSL, a component of the Notch signaling pathway, downregulated the expression of Penaeus vannamei hemocyanin small subunit gene (PvHMCs), indicating the involvement of
CSL in regulating the PvHMCs transcription. In this study, we identified a
CSL binding motif (\"GAATCCCAGA\", +1675/+1684 bp) in the core promoter of PvHMCs (designated as HsP3). Dual luciferase reporter assay and electrophoretic mobility shift assay (EMSA) demonstrated that the
CSL homolog in P. vannamei (PvCSL) could directly bind and activate the HsP3 promoter. Moreover, in vivo silencing of PvCSL significantly attenuated the mRNA and protein expression of PvHMCs. Finally, in response to Vibrio parahaemolyticus, Streptococcus iniae and white spot syndrome virus (WSSV) challenge, the transcript of PvCSL and PvHMCs showed a positive correlation, suggesting that PvCSL could also modulate the expression of PvHMCs upon pathogen stimulation. Taken together, our present finding is the first to demonstrate that PvCSL is a crucial factor in transcriptional control of PvHMCs.