Schistosoma is the causative agent of schistosomiasis, a common infectious disease distributed worldwide. Our previous phosphoproteomic analysis suggested that glycogen synthase kinase 3 (GSK3), a conserved protein kinase in eukaryotes, is likely involved in protein phosphorylation of Schistosoma japonicum. Here, we aimed to identify the interacting partners of S. japonicum GSK3β (SjGSK3β) and to evaluate its role in parasite survival. Toward these ends, we determined the transcription levels of SjGSK3β at different developmental stages and identified its interacting partners of SjGSK3β by screening a yeast two-hybrid S. japonicum cDNA library. We further used RNA interference (RNAi) to inhibit the expression of SjGSK3β in adult worms in vitro and examined the resultant changes in transcription of its putative interacting proteins and in worm viability compared with those of control worms. Reverse transcription-quantitative polymerase chain analysis indicated that SjGSK3β is expressed throughout the life cycle of S. japonicum, with higher expression levels detected in the eggs and relatively higher expression level found in male worms than in female worms. By screening the yeast two-hybrid library, eight proteins were identified as potentially interacting with SjGSK3β including cell division cycle 37 homolog (Cdc37), 14-3-3 protein, tegument antigen (I(H)A), V-ATPase proteolipid subunit, myosin alkali light chain 1, and three proteins without recognized functional domains. In addition, SjGSK3β RNAi reduced the SjGSK3β gene transcript level, leading to a significant decrease in kinase activity, cell viability, and worm survival. Collectively, these findings suggested that SjGSK3β may interact with its partner proteins to influence worm survival by regulating kinase activity.

血吸虫病是血吸虫病的病原体，血吸虫病是一种分布在世界各地的常见传染病。我们以前的磷酸化蛋白质组学分析表明，糖原合酶激酶3（GSK3）是真核生物中的保守蛋白激酶，可能与日本血吸虫的蛋白磷酸化有关。在这里，我们旨在确定日本血吸虫GSK3β（SjGSK3β）的相互作用伙伴，并评估其在寄生虫存活中的作用。为此，我们通过筛选酵母双杂交日本血吸虫cDNA文库确定了SjGSK3β在不同发育阶段的转录水平，并确定了其相互作用伴侣。我们进一步使用RNA干扰（RNAi）来抑制SjGSK3β在成虫体内进行了体外实验，并检查了与对照蠕虫相比，其推测的相互作用蛋白转录和蠕虫生存能力的变化。逆转录定量聚合酶链分析表明SjGSK3β在日本血吸虫的整个生命周期中都有表达，在卵中检测到较高的表达水平，而在雄性蠕虫中发现的表达水平要比雌性蠕虫中更高。通过筛选酵母双杂交文库，鉴定出八种蛋白可能与SjGSK3β相互作用，包括细胞分裂周期37同源物（Cdc37），14-3-3蛋白，外皮抗原（I（H）A），V-ATPase蛋白脂亚基，肌球蛋白碱轻链1和三种没有公认功能域的蛋白质。此外，SjGSK3βRNAi降低了SjGSK3β基因的转录水平，从而导致激酶活性，细胞活力和蠕虫存活率显着下降。这些发现共同表明，SjGSK3β可能与其伴侣蛋白相互作用，通过调节激酶活性来影响蠕虫的存活。