Jiangsu Key Laboratory of Neuropsychiatric Diseases and Department of Pharmacology, College of Pharmaceutical Sciences, Soochow University, Suzhou, China.
PLoS Biol. 2024 Jul 15;22(7):e3002716. doi: 10.1371/journal.pbio.3002716. eCollection 2024 Jul.
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.
腺苷酸环化酶（AC）的异源敏化导致 cAMP 信号转导升高，这有助于药物依赖。通过阻断 Cullin3 的 neddylation 来抑制 Cullin3-RING 连接酶可消除异源敏化，但是，调节机制仍不清楚。在这里，我们报告了钾通道四聚化结构域（KCTD）蛋白 2、5 和 17，特别是主要同工型 KCTD5 通过与 Cullin3 和 Cullin 相关和 neddylation 解离 1（CAND1）蛋白相互作用，在调节 AC1 的异源敏化和吗啡依赖中的重要作用。在细胞模型中，我们观察到 KCTD5 与 Gβ和 Cullin3 的结合增强，以及 Gβ与 AC1 的解离以及 CAND1 与 Cullin3 的解离增强，从而导致 AC1 的异源敏化。鉴于 CAND1 的结合抑制 Cullin3 的 neddylation，我们进一步阐明了 KCTD5 与 Gβ和 Cullin3 的相互作用增强促进了 CAND1 从 Cullin3 的解离，减弱了 CAND1 对 Cullin3 neddylation 的抑制作用，最终导致 AC1 的异源敏化。室旁丘脑核（PVT）在介导吗啡依赖中起重要作用。通过药理学和生化方法，我们随后证明 KCTD5/Cullin3 通过调节异源敏化的 AC（可能是小鼠 PVT 中的 AC1）来调节吗啡依赖。总之，本研究揭示了 Cullin3 介导的 AC1 异源敏化的潜在机制，并发现了 KCTD 蛋白在调节小鼠吗啡依赖中的作用。
