PMID- 30224479 OWN - NLM STAT- MEDLINE DCOM- 20181017 LR - 20211204 IS - 1091-6490 (Electronic) IS - 0027-8424 (Print) IS - 0027-8424 (Linking) VI - 115 IP - 40 DP - 2018 Oct 2 TI - Targeted profiling of RNA translation reveals mTOR-4EBP1/2-independent translation regulation of mRNAs encoding ribosomal proteins. PG - E9325-E9332 LID - 10.1073/pnas.1805782115 [doi] AB - The PI3K-Akt-mTOR signaling pathway is a master regulator of RNA translation. Pharmacological inhibition of this pathway preferentially and coordinately suppresses, in a 4EBP1/2-dependent manner, translation of mRNAs encoding ribosomal proteins. However, it is unclear whether mechanistic target of rapamycin (mTOR)-4EBP1/2 is the exclusive translation regulator of this group of genes, and furthermore, systematic searches for novel translation modulators have been immensely challenging because of difficulties in scaling existing RNA translation profiling assays. Here, we developed a rapid and highly scalable approach for gene-specific quantitation of RNA translation, termed Targeted Profiling of RNA Translation (TPRT). We applied this technique in a chemical screen for translation modulators, and identified numerous preclinical and clinical therapeutic compounds, with diverse nominal targets, that preferentially suppress translation of ribosomal proteins. Surprisingly, some of these compounds act in a manner that bypasses canonical regulation by mTOR-4EBP1/2. Instead, these compounds exert their translation effects in a manner that is dependent on GCN2-eIF2alpha, a central signaling axis within the integrated stress response. Furthermore, we were also able to identify metabolic perturbations that also suppress ribosomal protein translation in an mTOR-independent manner. Together, we describe a translation assay that is directly applicable to large-scale RNA translation studies, and that enabled us to identify a noncanonical, mTOR-independent mode for translation regulation of ribosomal proteins. FAU - Li, Ben B AU - Li BB AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. AD - Broad Institute of MIT and Harvard, Cambridge, MA 02142. FAU - Qian, Changli AU - Qian C AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. FAU - Gameiro, Paulo A AU - Gameiro PA AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. FAU - Liu, Chin-Chih AU - Liu CC AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. FAU - Jiang, Tao AU - Jiang T AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. FAU - Roberts, Thomas M AU - Roberts TM AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. FAU - Struhl, Kevin AU - Struhl K AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; kevin@hms.harvard.edu jean_zhao@dfci.harvard.edu. AD - Broad Institute of MIT and Harvard, Cambridge, MA 02142. FAU - Zhao, Jean J AU - Zhao JJ AD - Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215; kevin@hms.harvard.edu jean_zhao@dfci.harvard.edu. AD - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115. AD - Broad Institute of MIT and Harvard, Cambridge, MA 02142. LA - eng GR - R01 CA187918/CA/NCI NIH HHS/United States GR - P50 CA168504/CA/NCI NIH HHS/United States GR - R35 CA210057/CA/NCI NIH HHS/United States GR - R01 CA107486/CA/NCI NIH HHS/United States GR - UL1 TR001102/TR/NCATS NIH HHS/United States GR - P50 CA165962/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20180917 PL - United States TA - Proc Natl Acad Sci U S A JT - Proceedings of the National Academy of Sciences of the United States of America JID - 7505876 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Carrier Proteins) RN - 0 (EBNA1BP2 protein, human) RN - 0 (Eukaryotic Initiation Factor-2) RN - 0 (Multiprotein Complexes) RN - 0 (PA2G4 protein, human) RN - 0 (RNA, Messenger) RN - 0 (RNA-Binding Proteins) RN - 0 (Ribosomal Proteins) RN - EC 2.7.1.1 (MTOR protein, human) RN - EC 2.7.11.1 (EIF2AK4 protein, human) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Adaptor Proteins, Signal Transducing/genetics/*metabolism MH - Carrier Proteins/genetics/*metabolism MH - Cell Line, Transformed MH - Cell Line, Tumor MH - Eukaryotic Initiation Factor-2/genetics/metabolism MH - Humans MH - Multiprotein Complexes/genetics/*metabolism MH - *Protein Biosynthesis MH - Protein Serine-Threonine Kinases/genetics/metabolism MH - RNA, Messenger/genetics/*metabolism MH - RNA-Binding Proteins/genetics/*metabolism MH - Ribosomal Proteins/*biosynthesis/genetics MH - *Signal Transduction MH - TOR Serine-Threonine Kinases/genetics/*metabolism PMC - PMC6176620 OTO - NOTNLM OT - GCN2-eIF2alpha OT - mTOR OT - ribosomal proteins OT - translation control OT - translation profiling COIS- Conflict of interest statement: B.B.L. and J.J.Z. are coinventors on patent application PCT/US2017/039001. All other authors declare no conflict of interest. EDAT- 2018/09/19 06:00 MHDA- 2018/10/18 06:00 PMCR- 2019/04/02 CRDT- 2018/09/19 06:00 PHST- 2018/09/19 06:00 [pubmed] PHST- 2018/10/18 06:00 [medline] PHST- 2018/09/19 06:00 [entrez] PHST- 2019/04/02 00:00 [pmc-release] AID - 1805782115 [pii] AID - 201805782 [pii] AID - 10.1073/pnas.1805782115 [doi] PST - ppublish SO - Proc Natl Acad Sci U S A. 2018 Oct 2;115(40):E9325-E9332. doi: 10.1073/pnas.1805782115. Epub 2018 Sep 17.