腺苷酸环化酶(AC)的异源致敏导致cAMP信号转导升高,从而导致药物依赖。通过阻断cullin3的neddylation抑制cullin3-RING连接酶可以消除异源敏化,然而,调节机制仍然未知。这里,我们报道了钾通道四聚化结构域(KCTD)蛋白2,5和17,尤其是显性同工型KCTD5通过与cullin3和cullin相关和neddylation解离1(CAND1)蛋白的作用,在调节AC1和吗啡依赖的异源致敏中的重要作用.在细胞模型中,我们观察到KCTD5与Gβ和cullin3的增强关联,以及在AC1的异源致敏中Gβ与cullin3的解离升高。鉴于CAND1的结合抑制了cullin3的neddylation,我们进一步阐明了KCTD5与Gβ和cullin3的增强相互作用促进了CAND1与cullin3的解离,减弱了CAND1对cullin3neddylation的抑制作用,最终导致AC1的异源致敏。脑室旁丘脑核(PVT)在介导吗啡依赖中起重要作用。通过药理学和生物化学方法,然后我们证明KCTD5/cullin3通过调节AC的异源致敏调节吗啡依赖,可能AC1在小鼠PVT中。总之,本研究揭示了cullin3介导的AC1异源致敏的潜在机制,并发现了KCTD蛋白在调节小鼠吗啡依赖中的作用。
Heterologous sensitization of adenylyl cyclase (AC) results in elevated cAMP signaling transduction that contributes to drug dependence. Inhibiting cullin3-RING ligases by blocking the neddylation of cullin3 abolishes heterologous sensitization, however, the modulating mechanism remains uncharted. Here, we report an essential role of the potassium channel tetramerization domain (KCTD) protein 2, 5, and 17, especially the dominant isoform KCTD5 in regulating heterologous sensitization of AC1 and morphine dependence via working with cullin3 and the cullin-associated and neddylation-dissociated 1 (CAND1) protein. In cellular models, we observed enhanced association of KCTD5 with Gβ and cullin3, along with elevated dissociation of Gβ from AC1 as well as of CAND1 from cullin3 in heterologous sensitization of AC1. Given binding of CAND1 inhibits the neddylation of cullin3, we further elucidated that the enhanced interaction of KCTD5 with both Gβ and cullin3 promoted the dissociation of CAND1 from cullin3, attenuated the inhibitory effect of CAND1 on cullin3 neddylation, ultimately resulted in heterologous sensitization of AC1. The paraventricular thalamic nucleus (PVT) plays an important role in mediating morphine dependence. Through pharmacological and biochemical approaches, we then demonstrated that KCTD5/cullin3 regulates morphine dependence via modulating heterologous sensitization of AC, likely AC1 in PVT in mice. In summary, the present study revealed the underlying mechanism of heterologous sensitization of AC1 mediated by cullin3 and discovered the role of KCTD proteins in regulating morphine dependence in mice.