TY - JOUR
T1 - A novel rna synthesis inhibitor, stk160830, has negligible dna-intercalating activity for triggering a p53 response, and can inhibit p53-dependent apoptosis
AU - Morita, Akinori
AU - Ochi, Shintaro
AU - Satoh, Hidetoshi
AU - Ujita, Shohei
AU - Matsushita, Yosuke
AU - Tada, Kasumi
AU - Toyoda, Mihiro
AU - Nishiyama, Yuichi
AU - Mizuno, Kosuke
AU - Deguchi, Yuichi
AU - Suzuki, Keiji
AU - Tanaka, Yoshimasa
AU - Ueda, Hiroshi
AU - Inaba, Toshiya
AU - Hosoi, Yoshio
AU - Aoki, Shin
N1 - Funding Information:
Acknowledgments: This research was partially supported by the Platform Project for Supporting Drug Discovery and Life Science Research from AMED under Grant Number JP20am0101086 (support number 1030). This study was also supported by the Center for Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, and the Support Center for Advanced Medical Sciences, Tokushima University Graduate School of Biomedical Sciences.
Funding Information:
Funding: This study was partially supported by KAKENHI (19H03604 and 16K10396) from the Japan Society for the Promotion of Science.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/10
Y1 - 2021/10
N2 - RNA synthesis inhibitors and protein synthesis inhibitors are useful for investigating whether biological events with unknown mechanisms require transcription or translation; however, the dependence of RNA synthesis has been difficult to verify because many RNA synthesis inhibitors cause adverse events that trigger a p53 response. In this study, we screened a library containing 9600 core compounds and obtained STK160830 that shows anti-apoptotic effects in irradiated wild-type-p53-bearing human T-cell leukemia MOLT-4 cells and murine thymocytes. In many of the p53-impaired cells and p53-knockdown cells tested, STK160830 did not show a remarkable anti-apoptotic effect, suggesting that the anti-apoptotic activity is p53-dependent. In the expression analysis of p53, p53-target gene products, and reference proteins by immunoblotting, STK160830 down-regulated the expression of many of the proteins examined, and the downregulation correlated strongly with its inhibitory effect on cell death. mRNA expression analyses by qPCR and nascent RNA capture kit revealed that STK160830 showed a decreased mRNA expression, which was similar to that induced by the RNA synthesis inhibitor actinomycin D but differed to some extent. Furthermore, unlike other RNA synthesis inhibitors such as actinomycin D, p53 accumulation by STK160830 alone was negligible, and a DNA melting-curve analysis showed very weak DNA-intercalating activity, indicating that STK160830 is a useful inhibitor for RNA synthesis without triggering p53-mediated damage responses.
AB - RNA synthesis inhibitors and protein synthesis inhibitors are useful for investigating whether biological events with unknown mechanisms require transcription or translation; however, the dependence of RNA synthesis has been difficult to verify because many RNA synthesis inhibitors cause adverse events that trigger a p53 response. In this study, we screened a library containing 9600 core compounds and obtained STK160830 that shows anti-apoptotic effects in irradiated wild-type-p53-bearing human T-cell leukemia MOLT-4 cells and murine thymocytes. In many of the p53-impaired cells and p53-knockdown cells tested, STK160830 did not show a remarkable anti-apoptotic effect, suggesting that the anti-apoptotic activity is p53-dependent. In the expression analysis of p53, p53-target gene products, and reference proteins by immunoblotting, STK160830 down-regulated the expression of many of the proteins examined, and the downregulation correlated strongly with its inhibitory effect on cell death. mRNA expression analyses by qPCR and nascent RNA capture kit revealed that STK160830 showed a decreased mRNA expression, which was similar to that induced by the RNA synthesis inhibitor actinomycin D but differed to some extent. Furthermore, unlike other RNA synthesis inhibitors such as actinomycin D, p53 accumulation by STK160830 alone was negligible, and a DNA melting-curve analysis showed very weak DNA-intercalating activity, indicating that STK160830 is a useful inhibitor for RNA synthesis without triggering p53-mediated damage responses.
KW - Apoptosis
KW - DNA intercalator
KW - P53
KW - RNA synthesis inhibitor
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U2 - 10.3390/life11101087
DO - 10.3390/life11101087
M3 - Article
AN - SCOPUS:85118275619
VL - 11
JO - Life
JF - Life
SN - 0024-3019
IS - 10
M1 - 1087
ER -