Postsynaptic competition between calcineurin and PKA regulates mammalian sleep-wake cycles

• 成果类型: Article
• 署名作者: Wang, Yimeng; Cao, Siyu; Tone, Daisuke; Fujishima, Hiroshi; Yamada, Rikuhiro G.; Ohno, Rei-ichiro; Shi, Shoi; Matsuzawa, Kyoko; Yada, Saori; Kaneko, Mari; Sakamoto, Hirokazu; Onishi, Taichi; Ukai-Tadenuma, Maki; Ukai, Hideki; Hanashima, Carina; Hirose, Kenzo; Kiyonari, Hiroshi; Sumiyama, Kenta; Ode, Koji L.; Ueda, Hiroki R.
• 署名单位: University of Tokyo; RIKEN; Kurume University; RIKEN; University of Tokyo; Waseda University; RIKEN; University of Tsukuba; University of Tokyo; Nagoya University
• 刊物名称: Nature
• ISSN/ISSBN: 0028-4563
• DOI: 10.1038/s41586-024-08132-2
• 发表日期: 2024-12-12
• 页码: 412-+
• 关键词: protein-kinase-a; catalytic subunit; drosophila; mice; inhibition; cells; rna; phosphatase-1; transcription; generation

摘要

The phosphorylation of synaptic proteins is a significant biochemical reaction that controls the sleep-wake cycle in mammals(1-3). Protein phosphorylation in vivo is reversibly regulated by kinases and phosphatases. In this study, we investigate a pair of kinases and phosphatases that reciprocally regulate sleep duration. First, we perform a comprehensive screen of protein kinaseA (PKA) and phosphoprotein phosphatase (PPP) family genes by generating 40 gene knockout mouse lines using prenatal and postnatal CRISPR targeting. We identify a regulatory subunit of PKA (Prkar2b), a regulatory subunit of protein phosphatase1 (PP1; Pppr1r9b) and catalytic and regulatory subunits of calcineurin (also known as PP2B) (Ppp3ca and Ppp3r1) as sleep control genes. Using adeno-associated virus (AAV)-mediated stimulation of PKA and PP1-calcineurin activities, we show that PKA is a wake-promoting kinase, whereas PP1 and calcineurin function as sleep-promoting phosphatases. The importance of these phosphatases in sleep regulation is supported by the marked changes in sleep duration associated with their increased and decreased activities, ranging from approximately 17.3h per day (PP1 expression) to 4.3h per day (postnatal CRISPR targeting of calcineurin). Localization signals to the excitatory post-synapse are necessary for these phosphatases to exert their sleep-promoting effects. Furthermore, the wake-promoting effect of PKA localized to the excitatory post-synapse negated the sleep-promoting effect of PP1-calcineurin. These findings indicate that PKA and PP1-calcineurin have competing functions in sleep regulation at excitatory post-synapses.