Search bioRxiv⌕ Search

Biology subjects

Hatakeyama, R.

Publications and source records attributed to Hatakeyama, R..

2 recordsLinked to original sources

Substrate-specific regulation of mTORC1 activity by G protein-coupled receptors

The mammalian/mechanistic Target of Rapamycin Complex 1 (mTORC1) orchestrates cell growth and metabolism in response to diverse external cues. mTORC1 has a myriad of phosphorylation substrates, each playing important physiological roles. Emerging evidence suggests that mTORC1 can respond to upstream signals in a nuanced manner, enabling differential regulation of individual substrates and downstream biological processes. However, the nature of signals that determine the signaling selectivity of mTORC1 remains incompletely understood. Here, we studied mTORC1 regulation by G protein-coupled receptors (GPCRs). We found that phosphorylation of the Transcription Factor EB (TFEB), a non-canonical mTORC1 substrate that controls lysosome biogenesis, responds to GPCRs differently, compared to canonical mTORC1 substrates controlling protein synthesis such as S6K1 and 4EBP1. In particular, the muscarinic acetylcholine receptor M5 (M5R) promoted phosphorylation of S6K1 and 4EBP1, while triggering TFEB dephosphorylation. Consequently, M5R stimulated protein synthesis without inhibiting lysosome biogenesis. The regulations of an anabolic process and a catabolic process, albeit both governed by mTORC1, are thus decoupled downstream of M5R. This study highlights the importance of reassessing the effects of GPCRs on mTORC1 by concurrently monitoring individual substrates, a critical consideration to be made when evaluating GPCR ligands as therapeutic agents targeting the mTORC1 pathway.

cell biology↗

TORC1-containing signaling endosomes source membrane from vacuoles

Organelle biogenesis is fundamental to eukaryotic cell biology. Yeast signaling endosomes were recently identified as a signaling platform for the evolutionarily conserved Target of Rapamycin Complex 1 (TORC1) kinase complex. Despite the importance of signaling endosomes for TORC1-mediated control of cellular metabolism, how this organelle is generated has been a mystery. Here, we developed a system to induce synchronized de novo formation of signaling endosomes, enabling real-time monitoring of their biogenesis. Using this system, we identify vacuoles as a membrane source for newly formed signaling endosomes. Membrane supply from vacuoles is mediated by the CROP membrane-cutting complex, consisting of Atg18 PROPPIN and retromer subunits. The formation of signaling endosomes requires TORC1 activity, suggestive of a tightly regulated process. This study unveiled the first mechanistic principles and molecular participants of signaling endosome biogenesis. SummaryMembrane is supplied by vacuoles during biogenesis of yeast signaling endosomes, in a process mediated by the membrane-cutting CROP complex and promoted by TORC1 kinase activity.

cell biology↗