Phosphorylation of RNA Polymerase II by CDKC;2 Maintains the Arabidopsis Circadian Clock Period. (27th January 2022)

• Record Type: Journal Article
• Authors: Uehara, Takahiro N; Nonoyama, Takashi; Taki, Kyomi; Kuwata, Keiko; Sato, Ayato; Fujimoto, Kazuhiro J; Hirota, Tsuyoshi; Matsuo, Hiromi; Maeda, Akari E; Ono, Azusa; Takahara, Tomoaki T; Tsutsui, Hiroki; Suzuki, Takamasa; Yanai, Takeshi; Kay, Steve A; Itami, Kenichiro; Kinoshita, Toshinori; Yamaguchi, Junichiro; Nakamichi, Norihito
• Is Part Of: Plant & cell physiology. Volume 63:Number 4(2022)
• Page Start: 450
• Page End: 462
• Publication Date: 2022-01-27
• Subjects: Arabidopsis thaliana (Arabidopsis) -- CDKC;2 -- Chemical screening -- Circadian clock -- Pol II phosphorylation; Plant physiology -- Periodicals; Microbiology -- Periodicals; Cytology -- Periodicals; Cell Physiology -- Periodicals; Plant Physiological Phenomena -- Periodicals; Cytology; Microbiology; Plant physiology; Periodicals; 571.205
• DOI: 10.1093/pcp/pcac011
• Languages: English
• ISSNs: 0032-0781

Abstract

Abstract: The circadian clock is an internal timekeeping system that governs about 24 h biological rhythms of a broad range of developmental and metabolic activities. The clocks in eukaryotes are thought to rely on lineage-specific transcriptional–translational feedback loops. However, the mechanisms underlying the basic transcriptional regulation events for clock function have not yet been fully explored. Here, through a combination of chemical biology and genetic approaches, we demonstrate that phosphorylation of RNA polymerase II by CYCLIN DEPENDENT KINASE C; 2 (CDKC;2) is required for maintaining the circadian period in Arabidopsis. Chemical screening identified BML-259, the inhibitor of mammalian CDK2/CDK5, as a compound lengthening the circadian period of Arabidopsis. Short-term BML-259 treatment resulted in decreased expression of most clock-associated genes. Development of a chemical probe followed by affinity proteomics revealed that BML-259 binds to CDKC;2. Loss-of-function mutations of cdkc;2 caused a long period phenotype. In vitro experiments demonstrated that the CDKC;2 immunocomplex phosphorylates the C-terminal domain of RNA polymerase II, and BML-259 inhibits this phosphorylation. Collectively, this study suggests that transcriptional activity maintained by CDKC;2 is required for proper period length, which is an essential feature of the circadian clock in Arabidopsis.