State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, PR China; Guangdong Province Key Laboratory for Aquatic Economic Animals, Sun Yat-sen University, PR China.
College of Forestry and Landscape Architecture, South China Agriculture University, 510642, Guangzhou, PR China.
Dev Comp Immunol. 2020 May;106:103581. doi: 10.1016/j.dci.2019.103581. Epub 2019 Dec 17.
The NF-κB family is a set of evolutionarily conserved transcription factors that play central roles in various biological events. Dorsal is an invertebrate NF-κB family member that is essential for the regulation of immune responses. In the current study, the Dorsal gene from Scylla paramamosain (SpDorsal) was identified, which showed high homology to other crustacean Dorsal proteins. Expression of SpDorsal was highest in hemocytes and could be significantly changed after immune stimulations. In expression vector-transfected S2 cells, SpDorsal was mainly localized in the cytoplasm and could be efficiently translocated into the nucleus upon immune stimulations with the Gram-positive bacteria Staphylococcus aureus and poly (I:C), but not the Gram-negative bacteria Vibrio parahaemolyticus. As a transcription factor, SpDorsal could activate the promoter of S. paramamosain Hyastatin (SpHyastatin) in vitro, while S. paramamosain Cactus (SpCactus), a homolog of IκB, could interact with SpDorsal to prevent its nuclear translocation and inhibit its transcription factor activity. Silencing of SpDorsal in vivo using RNAi strategy significantly increased the mortality of crabs infected with S. aureus but not that with V. parahaemolyticus. These indicated that the SpDorsal signaling pathway could be mainly implicated in immune responses against Gram-positive bacterial infection in S. paramamosain.
NF-κB 家族是一组进化上保守的转录因子,在各种生物事件中发挥核心作用。Dorsal 是一种无脊椎动物 NF-κB 家族成员,对于免疫反应的调节至关重要。在本研究中,鉴定了锯缘青蟹(Scylla paramamosain)的 Dorsal 基因(SpDorsal),其与其他甲壳动物 Dorsal 蛋白具有高度同源性。SpDorsal 在血细胞中的表达最高,并且可以在免疫刺激后显著变化。在表达载体转染的 S2 细胞中,SpDorsal 主要定位于细胞质中,并且可以在受到革兰氏阳性细菌金黄色葡萄球菌和多聚(I:C)的免疫刺激后有效地易位到细胞核中,但不能在革兰氏阴性细菌副溶血弧菌的刺激下易位到细胞核中。作为一种转录因子,SpDorsal 可以在体外激活 S. paramamosain Hyastatin(SpHyastatin)的启动子,而 SpCactus,一种 IκB 的同源物,可以与 SpDorsal 相互作用,阻止其核易位并抑制其转录因子活性。使用 RNAi 策略在体内沉默 SpDorsal 会显著增加感染金黄色葡萄球菌的螃蟹的死亡率,但对感染副溶血弧菌的螃蟹的死亡率没有影响。这些表明 SpDorsal 信号通路可能主要参与锯缘青蟹对革兰氏阳性细菌感染的免疫反应。