Maintaining ATP levels via the suppression of PERK-mediated rRNA synthesis at ER stress.
Currently, [(3)H]uridine is most often used to monitor rRNA synthesis in cultured cells. We show here that radiolabeled ribonucleoside triphosphates, such as [alpha-(33)P]UTP, in culture medium were also incorporated efficiently not only into cells but also into de novo RNA, particularly rRNA. Using this method, we first revealed that endoplasmic reticulum (ER) stress inducers such as tunicamycin and thapsigargin suppressed de novo rRNA synthesis, and that PERK, but not IRE1alpha or ATF6, mediated the suppression. PERK is known to mediate the suppression of de novo protein synthesis via phosphorylation of eIF2alpha. Consistently, other translational inhibitors such as PSI, proteasomal inhibitor, and cycloheximide suppressed de novo rRNA synthesis. eIF2alpha knockdown also suppressed both de novo protein and rRNA syntheses. Furthermore, ER stress reduced cellular ATP levels, and the suppression of rRNA synthesis apparently mitigated their reduction. These observations provided a close link between ATP levels and suppression of de novo rRNA synthesis at ER stress, and we proposed a novel feedback mechanism, in which ATP levels were maintained via suppression of de novo rRNA synthesis in ATP-demanding stresses, such as ER stress.
Okamoto
Akihiko
A
The Laboratory of Functional Biology, Kyoto University Graduate School of Biostudies/SORST (JST), Kyoto 606-8501, Japan.
Koike
Masaaki
M
Yasuda
Kunihiko
K
Kakizuka
Akira
A
D000255:Adenosine Triphosphate / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D006367:HeLa Cells; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D012335:RNA, Ribosomal / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D013312:Stress, Physiological; D019284:Thapsigargin / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology; D019892:eIF-2 Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000255:Adenosine Triphosphate; D047428:Protein Kinase Inhibitors; D012335:RNA, Ribosomal; D019284:Thapsigargin; D014415:Tunicamycin; D019892:eIF-2 Kinase