Moleclar genetic analysis of eukaryotic molybdate transport systems in a model yeast Hansenula polymorpha and a model animal cell

钼是几乎所有生物必需的微量元素。虽然已知活细胞从周围环境输入无机钼酸根离子，但细胞内钼酸根离子和/或转运系统的体内动力学仍然知之甚少。在本研究中，我们研究了一种模型单细胞酵母多形汉逊酵母的钼酸盐转运系统。遗传分析结果表明，该酵母菌至少具有三种不同的钼酸盐运输类型。此外，我们还获得了三个钼酸盐运输的候选基因，它们在异源表达系统中表现出钼酸根积累。此外，我们开发了一种基于遗传编码的Forester-Resonance-Energy Transfer(FRET)的钼酸根纳米传感器。利用该纳米传感器，我们在模型动物细胞HEK293T中展示了钼酸根的活体动力学。动力学分析表明，新型草酸敏感和硫酸盐抗性转运蛋白(S)在细胞中摄取钼酸盐。

Molybdenum (Mo) is an essential trace element for almost all living organisms. Whilst living cells are known to import inorganic molybdate oxyanion from the surrounding environment, the in vivo dynamics of cytosolic molybdate and/or transport systems remain poorly understood. We investigated molybdate transport system of a model unicellular yeast Hansenula polymorpha in this study. Results of genetic analysis suggested the yeast has at least three defferent type of molybdate transport. Also, we obtained three candidate genes for the molybdate transport those exhibited molybdate accumulation in heterologous expression systems. Furthermore, we developed a genetically encoded Forester-resonance-energy-transfer (FRET)-based nanosensor for molybdate. By using the nanosensor we demonstrated in vivo dynamics of molybdate in a model animal cell HEK293T . Analyses of the dynamics suggest that novel oxalate-sensitive- and sulfate-resistant- transporter(s) uptake molybdate in thecell.