Silva Gunawardene Y I, Chow B K, He J G, Chan S M
Department of Zoology, The University of Hong Kong, Pokfulam Road, Hong Kong, People's Republic of China.
Insect Biochem Mol Biol. 2001 Oct;31(11):1115-24. doi: 10.1016/s0965-1748(01)00060-1.
Methylfarnesoate (MF), an analogue of the insect juvenile hormone III, has been implicated to play a vital role in the regulation of the growth and reproductive development in crustaceans. Farnesoic acid O-methyltransferase (FAMeT) is the key enzyme involved in catalyzing the final step in the MF biosynthetic pathway. In this study, we report the cloning and characterization of the cDNA encoding the putative FAMeT of the shrimp Metapenaeus ensis. FAMeT comprises 280 amino acid residues with a predicted molecular weight of 32kDa. The predicted putative FAMeT protein reveals a high degree of structural conservation of FAMeT with the lobsters. It shares 79 and 70% sequence identities with the putative FAMeTs of Homarus americanus and Panulirus interruptus, respectively. As revealed by the Southern blot analysis and genomic PCR, only one gene exists in the shrimp genome and the gene is uninterrupted in the coding region. The shrimp FAMeT mRNA is widely distributed in many tissues with the highest expression level observed in the central nervous system. A constant level of FAMeT expression is recorded in the ventral nerve cord of the juveniles and the mature females during the reproductive cycle. Unlike the ventral nerve cord, the eyestalk of the juvenile male, but not the female, expresses FAMeT. Further study shows that the eyestalk of the mature female expresses FAMeT during all stages of ovarian maturation. We speculate that FAMeT may be important for the regulation of eyestalk neuropeptides. This is the first extensive study on the molecular characterization, structural analysis and expression of the crustacean FAMeT.
法尼酸甲酯(MF)是昆虫保幼激素III的类似物,被认为在甲壳类动物的生长和生殖发育调节中起着至关重要的作用。法尼酸O - 甲基转移酶(FAMeT)是催化MF生物合成途径最后一步的关键酶。在本研究中,我们报告了对虾新对虾假定的FAMeT编码cDNA的克隆和表征。FAMeT由280个氨基酸残基组成,预测分子量为32kDa。预测的假定FAMeT蛋白与龙虾的FAMeT具有高度的结构保守性。它与美洲螯龙虾和断沟龙虾假定的FAMeT分别具有79%和70%的序列同一性。Southern印迹分析和基因组PCR结果显示,对虾基因组中仅存在一个基因,且该基因在编码区无中断。对虾FAMeT mRNA广泛分布于许多组织中,在中枢神经系统中表达水平最高。在幼体和成熟雌性的生殖周期中,腹神经索中FAMeT的表达水平保持恒定。与腹神经索不同,幼体雄性而非雌性的眼柄表达FAMeT。进一步研究表明,成熟雌性的眼柄在卵巢成熟的所有阶段均表达FAMeT。我们推测FAMeT可能对眼柄神经肽的调节很重要。这是首次对甲壳类动物FAMeT进行的关于分子表征、结构分析和表达的广泛研究。