首页> 美国卫生研究院文献>The Journal of Biological Chemistry >Heat Shock Protein 83 (Hsp83) Facilitates Methoprene-tolerant (Met) Nuclear Import to Modulate Juvenile Hormone Signaling
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Heat Shock Protein 83 (Hsp83) Facilitates Methoprene-tolerant (Met) Nuclear Import to Modulate Juvenile Hormone Signaling

机译:热休克蛋白83(Hsp83)促进甲氧戊丁耐受(Met)核输入以调节少年激素信号转导。

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摘要

Juvenile hormone (JH) receptors, methoprene-tolerant (Met) and Germ-cell expressed (Gce), transduce JH signals to induce Kr-h1 expression in Drosophila. Dual luciferase assay identified a 120-bp JH response region (JHRR) in the Kr-h1α promoter. Both in vitro and in vivo experiments revealed that Met and Gce transduce JH signals to induce Kr-h1 expression through the JHRR. DNA affinity purification identified chaperone protein Hsp83 as one of the proteins bound to the JHRR in the presence of JH. Interestingly, Hsp83 physically interacts with PAS-B and basic helix-loop-helix domains of Met, and JH induces Met-Hsp83 interaction. As determined by immunohistochemistry, Met is mainly distributed in the cytoplasm of fat body cells of the larval when the JH titer is low and JH induces Met nuclear import. Hsp83 was accumulated in the cytoplasm area adjunct to the nucleus in the presence of JH and Met/Gce. Loss-of-function of Hsp83 attenuated JH binding and JH-induced nuclear import of Met, resulting in a decrease in the JHRR-driven reporter activity leading to reduction of Kr-h1 expression. These data show that Hsp83 facilitates the JH-induced nuclear import of Met that induces Kr-h1 expression through the JHRR.
机译:甲氧戊二烯耐受(Met)和生殖细胞表达(Gce)的少年激素(JH)受体转导JH信号以诱导果蝇中Kr-h1表达。双重荧光素酶测定法在Kr-h1α启动子中鉴定出一个120 bp的JH反应区(JHRR)。体外和体内实验均表明,Met和Gce转导JH信号以通过JHRR诱导Kr-h1表达。 DNA亲和纯化将伴侣蛋白Hsp83鉴定为在JH存在下与JHRR结合的蛋白之一。有趣的是,Hsp83与Met的PAS-B和基本螺旋-环-螺旋结构域发生物理相互作用,而JH诱导Met-Hsp83相互作用。通过免疫组织化学测定,当JH滴度低且JH诱导Met核输入时,Met主要分布在幼虫的脂肪体细胞的细胞质中。在JH和Met / Gce的存在下,Hsp83积累在细胞核附近的细胞质区域。 Hsp83的功能丧失减弱了JH结合和JH诱导的Met核输入,导致JHRR驱动的报道分子活性降低,导致Kr-h1表达降低。这些数据表明,Hsp83促进了JH诱导的Met核输入,从而通过JHRR诱导Kr-h1表达。

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