TY - JOUR
T1 - Identification of novel compounds that inhibit SnRK2 kinase activity by high-throughput screening
AU - Matsuoka, Shoko
AU - Sato, Karin
AU - Maruki-Imamura, Riyo
AU - Noutoshi, Yoshiteru
AU - Okabe, Takayoshi
AU - Kojima, Hirotatsu
AU - Umezawa, Taishi
N1 - Funding Information:
We thank Dr. Jeffrey C. Anderson (OSU, USA) for comments about and proofreading of this manuscript. We also thank Ryo Yanagisawa for discussions about this study and Izumi Minegishi and other DDI staff for preparation of the plates containing the compounds. This research was supported by the Platform Project for Supporting Drug Discovery and Life Science Research from AMED under Grant Number JP19am0101086 (support number 1069). T87 suspension cells were provided by RIKEN BRC. This work was partly supported by JST PRESTO [P13413773] to T.U.
Funding Information:
We thank Dr. Jeffrey C. Anderson (OSU, USA) for comments about and proofreading of this manuscript. We also thank Ryo Yanagisawa for discussions about this study and Izumi Minegishi and other DDI staff for preparation of the plates containing the compounds. This research was supported by the Platform Project for Supporting Drug Discovery and Life Science Research from AMED under Grant Number JP19am0101086 (support number 1069 ). T87 suspension cells were provided by RIKEN BRC. This work was partly supported by JST PRESTO [P13413773] to T.U.
Publisher Copyright:
© 2020
PY - 2021/1/22
Y1 - 2021/1/22
N2 - Abscisic acid (ABA) is a major phytohormone that regulates abiotic stress responses and development. SNF1-rerated protein kinase 2 (SnRK2) is a key regulator of ABA signaling. To isolate compounds which directly affect SnRK2 activity, we optimized a fluorescence-based system for high-throughput screening (HTS) of SnRK2 kinase regulators. Using this system, we screened a chemical library consisting of 16,000 compounds and identified ten compounds (INH1-10) as potential SnRK2 inhibitors. Further characterization of these compounds by in vitro phosphorylation assays confirmed that three of the ten compounds were SnRK2-specific kinase inhibitors. In contrast, seven of ten compounds inhibited ABA-responsive gene expression in Arabidopsis cells. From these results, INH1 was identified as a SnRK2-specific inhibitor in vitro and in vivo. We propose that INH1 could be a lead compound of chemical tools for studying ABA responses in various plant species.
AB - Abscisic acid (ABA) is a major phytohormone that regulates abiotic stress responses and development. SNF1-rerated protein kinase 2 (SnRK2) is a key regulator of ABA signaling. To isolate compounds which directly affect SnRK2 activity, we optimized a fluorescence-based system for high-throughput screening (HTS) of SnRK2 kinase regulators. Using this system, we screened a chemical library consisting of 16,000 compounds and identified ten compounds (INH1-10) as potential SnRK2 inhibitors. Further characterization of these compounds by in vitro phosphorylation assays confirmed that three of the ten compounds were SnRK2-specific kinase inhibitors. In contrast, seven of ten compounds inhibited ABA-responsive gene expression in Arabidopsis cells. From these results, INH1 was identified as a SnRK2-specific inhibitor in vitro and in vivo. We propose that INH1 could be a lead compound of chemical tools for studying ABA responses in various plant species.
KW - ABA signaling
KW - Drought stress
KW - High-throughput screening
KW - Kinase inhibitor
KW - SNF1-Related protein kinase 2
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U2 - 10.1016/j.bbrc.2020.12.046
DO - 10.1016/j.bbrc.2020.12.046
M3 - Article
C2 - 33385806
AN - SCOPUS:85098546754
SN - 0006-291X
VL - 537
SP - 57
EP - 63
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
ER -