Mita M, Yoshikuni M, Nagahama Y
Department of Biology, School of Education, Waseda University, Tokyo 169-8050, Japan.
Biochim Biophys Acta. 1999 Jun 28;1428(1):13-20. doi: 10.1016/s0304-4165(99)00041-0.
1-Methyladenine (1-MeAde), the oocyte maturation-inducing substance in starfish, is produced by ovarian follicle cells upon stimulation with a gonad-stimulating substance (GSS) released from the radial nerves. We have shown previously that GSS causes a reduction in the intracellular levels of ATP coincident with 1-MeAde production. The present study examined whether the adenine molecule of 1-MeAde is directly derived from ATP. When isolated follicle cells from the starfish Asterina pectinifera were preloaded with [U-14C]adenine or [U-14C]adenosine, there was an increase in the intracellular levels of radiolabeled adenine nucleotides, particularly ATP. Following further incubation with GSS, the intracellular levels of radiolabeled ATP decreased, concomitant with a marked increase in the levels of [14C]1-MeAde in the medium. The amount of ATP consumed under the influence of GSS was similar to the amount of 1-MeAde produced. However, there was no change in the levels of ADP and AMP regardless of the presence or absence of GSS. These findings strongly suggest that 1-MeAde is synthesized from ATP as a substrate in follicle cells under the influence of GSS. Furthermore, using [methyl-3H]methionine, the methyl group of 1-MeAde was found to be derived from methionine. Thus GSS appears to stimulate the synthesis of 1-MeAde from ATP via the methylation process in starfish ovarian follicle cells.
1-甲基腺嘌呤(1-MeAde)是海星中诱导卵母细胞成熟的物质,由卵巢滤泡细胞在受到从放射神经释放的促性腺物质(GSS)刺激后产生。我们之前已经表明,GSS会导致细胞内ATP水平降低,同时产生1-MeAde。本研究检测了1-MeAde的腺嘌呤分子是否直接来源于ATP。当从多棘海盘车分离的滤泡细胞预先加载[U-14C]腺嘌呤或[U-14C]腺苷时,细胞内放射性标记的腺嘌呤核苷酸水平增加,尤其是ATP。在用GSS进一步孵育后,细胞内放射性标记的ATP水平降低,同时培养基中[14C]1-MeAde的水平显著增加。在GSS影响下消耗的ATP量与产生的1-MeAde量相似。然而,无论是否存在GSS,ADP和AMP的水平都没有变化。这些发现强烈表明,在GSS的影响下,1-MeAde在滤泡细胞中以ATP为底物合成。此外,使用[甲基-3H]甲硫氨酸,发现1-MeAde的甲基来源于甲硫氨酸。因此,GSS似乎通过海星卵巢滤泡细胞中的甲基化过程刺激从ATP合成1-MeAde。