Niwa Ryusuke, Niimi Teruyuki, Honda Naoko, Yoshiyama Michiyo, Itoyama Kyo, Kataoka Hiroshi, Shinoda Tetsuro
Initiative for the Promotion of Young Scientists' Independent Research, Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan.
Insect Biochem Mol Biol. 2008 Jul;38(7):714-20. doi: 10.1016/j.ibmb.2008.04.003. Epub 2008 Apr 30.
Juvenile hormone (JH) acid O-methyltransferase (JHAMT) is the enzyme that transfers a methyl group from S-adenosyl-l-methionine (SAM) to the carboxyl group of JH acids to produce active JHs in the corpora allata. While the JHAMT gene was originally identified and characterized in the silkworm Bombyx mori, no orthologs from other insects have been studied until now. Here we report on the functional characterization of the CG17330/DmJHAMT gene in the fruit fly Drosophila melanogaster. Recombinant DmJHAMT protein expressed in Escherichia coli catalyzes the conversion of farnesoic acid and JH III acid to their cognate methyl esters in the presence of SAM. DmJHAMT is predominantly expressed in corpora allata, and its developmental expression profile correlates with changes in the JH titer. While a transgenic RNA interference against DmJHAMT has no visible effect, overexpression of DmJHAMT results in a pharate adult lethal phenotype, similar to that obtained with application of JH analogs, suggesting that the temporal regulation of DmJHAMT is critical for Drosophila development.
保幼激素(JH)酸O-甲基转移酶(JHAMT)是一种将S-腺苷-L-甲硫氨酸(SAM)上的甲基转移至JH酸羧基上,从而在咽侧体中产生活性JH的酶。虽然JHAMT基因最初是在家蚕中被鉴定和表征的,但直到现在还没有对其他昆虫的直系同源基因进行过研究。在此,我们报道了果蝇黑腹果蝇中CG17330/DmJHAMT基因的功能特性。在大肠杆菌中表达的重组DmJHAMT蛋白在SAM存在的情况下,催化法尼酸和JH III酸转化为它们相应的甲酯。DmJHAMT主要在咽侧体中表达,其发育表达谱与JH滴度的变化相关。虽然针对DmJHAMT的转基因RNA干扰没有明显影响,但DmJHAMT的过表达会导致蛹期成虫致死表型,类似于使用JH类似物所获得的表型,这表明DmJHAMT的时间调控对果蝇发育至关重要。