TY  - JOUR
AU  - Hermann, Julia
AU  - Bortecen, Toman
AU  - Kalis, Robert
AU  - Kowar, Alexander
AU  - Pechincha, Catarina
AU  - Vogt, Vivien
AU  - Schneider, Martin
AU  - Helm, Dominic
AU  - Krijgsveld, Jeroen
AU  - Loayza-Puch, Fabricio
AU  - Zuber, Johannes
AU  - Palm, Wilhelm
TI  - mTORC1 cooperates with tRNA wobble modification to sustain the protein synthesis machinery.
JO  - Nature Communications
VL  - 16
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Springer Nature
M1  - DKFZ-2025-00942
SP  - 4201
PY  - 2025
N1  - DKFZ-ZMBH Alliance / #EA:A330#LA:A330#
AB  - Synthesizing the cellular proteome is a demanding process that is regulated by numerous signaling pathways and RNA modifications. How precisely these mechanisms control the protein synthesis machinery to generate specific proteome subsets remains unclear. Here, through genome-wide CRISPR screens we identify genes that enable mammalian cells to adapt to inactivation of the kinase mechanistic target of rapamycin complex 1 (mTORC1), the central driver of protein synthesis. When mTORC1 is inactive, enzymes that modify tRNAs at wobble uridines (U34-enzymes), Elongator and Ctu1/2, become critically essential for cell growth in vitro and in tumors. By integrating quantitative nascent proteomics, steady-state proteomics and ribosome profiling, we demonstrate that the loss of U34-enzymes particularly impairs the synthesis of ribosomal proteins. However, when mTORC1 is active, this biosynthetic defect only mildly affects steady-state protein abundance. By contrast, simultaneous suppression of mTORC1 and U34-enzymes depletes cells of ribosomal proteins, globally inhibiting translation. Thus, mTORC1 cooperates with tRNA U34-enzymes to sustain the protein synthesis machinery and support the high translational requirements of cell growth.
KW  - Mechanistic Target of Rapamycin Complex 1: metabolism
KW  - Mechanistic Target of Rapamycin Complex 1: genetics
KW  - RNA, Transfer: metabolism
KW  - RNA, Transfer: genetics
KW  - Protein Biosynthesis
KW  - Humans
KW  - Ribosomes: metabolism
KW  - Ribosomal Proteins: metabolism
KW  - Ribosomal Proteins: genetics
KW  - HEK293 Cells
KW  - Animals
KW  - Proteomics
KW  - Uridine: metabolism
KW  - Mechanistic Target of Rapamycin Complex 1 (NLM Chemicals)
KW  - RNA, Transfer (NLM Chemicals)
KW  - Ribosomal Proteins (NLM Chemicals)
KW  - Uridine (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:40328729
C2  - pmc:PMC12056009
DO  - DOI:10.1038/s41467-025-59185-4
UR  - https://inrepo02.dkfz.de/record/300828
ER  -