*Key Laboratory of Conservation Biology for Endangered Wildlife of the Ministry of Education and College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou 310058, Zhejiang, People's Republic of China.
Biochem J. 2013 Dec 1;456(2):185-94. doi: 10.1042/BJ20131020.
Trehalase, which specifically hydrolyses trehalose into glucose, plays an important role in the metabolism of trehalose. Large amounts of trehalose are stored in the diapause encysted embryos (cysts) of Artemia, which are not only vital to their extraordinary stress resistance, but also provide a source of energy for development after diapause is terminated. In the present study, a mechanism for the transcriptional regulation of trehalase was described in Artemia parthenogenetica. A trehalase-associated protein (ArTAP) was identified in Artemia-producing diapause cysts. ArTAP was found to be expressed only in diapause-destined embryos. Further analyses revealed that ArTAP can bind to a specific intronic segment of a trehalase gene. Knockdown of ArTAP by RNAi resulted in the release of cysts with coarse shells in which two chitin-binding proteins were missing. Western blotting showed that the level of trehalase was increased and apoptosis was induced in these ArTAP-knockdown cysts compared with controls. Taken together, these results show that ArTAP is a key regulator of trehalase expression which, in turn, plays an important role in trehalose metabolism during the formation of diapause cysts.
海藻糖酶特异性地将海藻糖水解为葡萄糖,在海藻糖代谢中发挥重要作用。大量的海藻糖储存在卤虫休眠包囊胚胎(休眠囊)中,这不仅对其非凡的抗逆性至关重要,而且为休眠结束后的发育提供了能量来源。本研究描述了卤虫孤雌生殖中海藻糖酶转录调控的机制。在产生休眠的卤虫休眠囊中产生活性的海藻糖酶相关蛋白(ArTAP)。发现 ArTAP 仅在休眠定向胚胎中表达。进一步的分析表明,ArTAP 可以与海藻糖酶基因的特定内含子片段结合。通过 RNAi 敲低 ArTAP 导致休眠囊释放,其中缺失了两种几丁质结合蛋白。Western blot 显示,与对照组相比,这些 ArTAP 敲低的休眠囊中海藻糖酶的水平增加,并且诱导了细胞凋亡。综上所述,这些结果表明 ArTAP 是海藻糖酶表达的关键调节剂,而海藻糖酶在休眠囊形成过程中的海藻糖代谢中发挥重要作用。