Molthathong Sudkhate, Senapin Saengchan, Klinbunga Sirawut, Puanglarp Narongsak, Rojtinnakorn Jiraporn, Flegel Timothy W
Centex Shrimp, Faculty of Science, Mahidol University, Bangkok, Thailand.
Fish Shellfish Immunol. 2008 Feb;24(2):173-9. doi: 10.1016/j.fsi.2007.10.013. Epub 2007 Nov 5.
Here we describe the highly conserved gene, defender against apoptotic death (DAD1) identified from an EST library of the black tiger shrimp Penaeus monodon. The full-length cDNA of DAD1 of P. monodon comprised 638bp with an ORF of 345bp corresponding to 114 deduced amino acids. The deduced amino acid sequence was compared to known DAD1 sequences in the GenBank and in other databases. Phylogenetic analysis revealed that P. monodon DAD1 clustered with DAD1 from other invertebrates. Real-time RT-PCR with RNA extracts from normal P. monodon revealed DAD1 expression in several tissues including those of digestive and defense organs such as the hepatopancreas and hemocytes, respectively. If death from YHV infection was related to increased levels of apoptosis, we reasoned that the level of DAD1 should decrease as YHV infections progressed, especially in hemocytes (HC), one of its main targets. Real-time RT-PCR with RNA extracts from HC of P. monodon challenged with YHV revealed that the transcriptional level of DAD1 declined dramatically (approximately 50%) after YHV challenge. Although this suggests that DAD1 plays a role in mortality caused by YHV, control of apoptosis is complex and involves the interaction of many proteins, few of which have been characterized for shrimp. Thus, firm conclusions regarding the role of DAD1 must await the description and characterization of other proteins.
在此,我们描述了从黑虎虾(斑节对虾)的一个EST文库中鉴定出的高度保守基因——抗凋亡死亡因子(DAD1)。斑节对虾DAD1的全长cDNA由638bp组成,其开放阅读框(ORF)为345bp,对应114个推导氨基酸。将推导的氨基酸序列与GenBank及其他数据库中的已知DAD1序列进行了比较。系统发育分析表明,斑节对虾DAD1与其他无脊椎动物的DAD1聚集在一起。对正常斑节对虾的RNA提取物进行实时RT-PCR分析,结果显示DAD1在包括消化器官(如肝胰腺)和防御器官(如血细胞)等多个组织中均有表达。如果黄头病毒(YHV)感染导致的死亡与凋亡水平升高有关,我们推测随着YHV感染的进展,DAD1的水平应会下降,尤其是在其主要靶标之一的血细胞(HC)中。对受YHV攻击的斑节对虾血细胞的RNA提取物进行实时RT-PCR分析,结果显示YHV攻击后DAD1的转录水平急剧下降(约50%)。虽然这表明DAD1在YHV引起的死亡率中发挥作用,但细胞凋亡的调控是复杂的,涉及许多蛋白质的相互作用,而其中很少有蛋白在对虾中得到表征。因此,关于DAD1作用的确切结论必须等待对其他蛋白质的描述和表征。