RNA editing enzyme ADAR1 governs the circadian expression of P-glycoprotein in human renal cells by regulating alternative splicing of the ABCB1 gene
| Autor: | Yuji Omata, Satoru Koyanagi, Tomoaki Yamauchi, Shigehiro Ohdo, Naoya Matsunaga, Akito Tsuruta |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
circadian rhythm
0301 basic medicine RNA editing ATF4 activating transcription factor 4 SLC solute carrier Adenosine Deaminase Circadian clock posttranscriptional regulation DEX dexamethasone Biology Kidney Biochemistry Cell Line alternative splicing 03 medical and health sciences P-gp P-glycoprotein HLF hepatic leukemia factor Humans shRNA small hairpin RNA ATP Binding Cassette Transporter Subfamily B Member 1 A-to-I adenosine-to-inosine Molecular Biology KD knockdown Gene knockdown Messenger RNA NMD nonsense-mediated mRNA decay SCN suprachiasmatic nucleus 030102 biochemistry & molecular biology Alternative splicing snRNP small nuclear ribonucleoprotein Intron RNA-Binding Proteins RNA RIP RNA immunoprecipitation Cell Biology Cell biology RNA silencing 030104 developmental biology RPTEC renal proximal tubular epithelial cell Gene Expression Regulation AhR aryl hydrocarbon receptor dsRNA double-stranded RNA ABC transporter ADAR adenosine deaminase acting on RNA pharmacokinetics Research Article |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/j.jbc.2021.100601 |
| Popis: | The expression and function of some xenobiotic transporters vary according to the time of the day, causing the dosing time-dependent changes in drug disposition and toxicity. P-glycoprotein (P-gp), encoded by the ABCB1 gene, is highly expressed in the kidneys and functions in the renal elimination of various drugs. The elimination of several P-gp substrates was demonstrated to vary depending on administration time, but the underlying mechanism remains unclear. We found that adenosine deaminase acting on RNA (ADAR1) was involved in the circadian regulation of P-gp expression in human renal proximal tubular epithelial cells (RPTECs). After synchronization of the cellular circadian clock by dexamethasone treatment, the expression of P-gp exhibited a significant 24-h oscillation in RPTECs, but this oscillation was disrupted by the downregulation of ADAR1. Although ADAR1 catalyzes adenosine-to-inosine (A-to-I) RNA editing in double-stranded RNA substrates, no significant ADAR1-regulated editing sites were detected in the human ABCB1 transcripts in RPTECs. On the other hand, downregulation of ADAR1 induced alternative splicing in intron 27 of the human ABCB1 gene, resulting in the production of retained intron transcripts. The aberrant spliced transcript was sensitive to nonsense-mediated mRNA decay, leading to the decreased stability of ABCB1 mRNA and prevention of the 24-h oscillation of P-gp expression. These findings support the notion that ADAR1-mediated regulation of alternative splicing of the ABCB1 gene is a key mechanism of circadian expression of P-gp in RPTECs, and the regulatory mechanism may underlie the dosing time-dependent variations in the renal elimination of P-gp substrates. |
| Databáze: | OpenAIRE |
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