Only in Titles

Search results for: Biotin

paperclip

#35973513   2022/08/13 To Up

Proximity labeling of endogenous RICTOR identifies mTOR Complex 2 regulation by ADP ribosylation factor ARF1.

Mechanistic Target of Rapamycin (mTOR) complex 2 (mTORC2) regulates metabolism, cell proliferation, and cell survival. mTORC2 activity is stimulated by growth factors, and it phosphorylates the hydrophobic motif site of the AGC kinases AKT, SGK, and PKC. However, the proteins that interact with mTORC2 to control its activity and localization remain poorly defined. To identify mTORC2 interacting proteins in living cells, we tagged endogenous RICTOR, an essential mTORC2 subunit, with the modified BirA biotin ligase BioID2 and performed live-cell proximity labeling. We identified 215 RICTOR-proximal proteins, including proteins with known mTORC2 pathway interactions, and 135 proteins (63%) not previously linked to mTORC2 signaling, including nuclear and cytoplasmic proteins. Our imaging and cell fractionation experiments suggest nearly 30% of RICTOR is in the nucleus, hinting at potential nuclear functions. We also identified 29 interactors containing RICTOR-dependent, insulin-stimulated phosphorylation sites, thus providing insight into mTORC2-dependent insulin signaling dynamics. Finally, we identify the endogenous ADP ribosylation factor 1 (ARF1) GTPase as an mTORC2-interacting protein. Through gain- and loss-of-function studies, we provide functional evidence that ARF1 may negatively regulate mTORC2. In summary, we present a new method of studying endogenous mTORC2, a resource of RICTOR/mTORC2 protein interactions in living cells, and a potential mechanism of mTORC2 regulation by the ARF1 GTPase.
Amelia K Luciano, Ekaterina Korobkina, Scott P Lyons, John A Haley, Shelagh Fluharty, Su Myung Jung, Arminja N Kettenbach, David A Guertin

1651 related Products with: Proximity labeling of endogenous RICTOR identifies mTOR Complex 2 regulation by ADP ribosylation factor ARF1.

200ul250ul100ug250ul5mg200ug5 x 50 ug200ul250ul100ul50 ug 250ul

Related Pathways

paperclip

#35973279   2022/08/11 To Up

A light-controlled DNA nanothermometer for temperature sensing in the cellular membrane microenvironment.

Precise sensing of cellular temperature is one significant yet challenge task for studying miscellaneous biological processes. Herein, we report a light-controlled DNA nanothermometer that allow for real-time thermal sensing in extracellular microscope with high spatiotemporal resolution. The light-controlled DNA nanothermometer three key elements: a thermal-sensitive molecular beacon (MB) labelled with fluorophore Cy5 and Cy3 at its 5' and 3' termini, an inhibitor strand containing two photocleavable linkers (pc-linker), and a biotin modified strand, which could modify this three-strand hybridization complex onto the cell surface. Upon exposing to UV light irradiation, the light-controlled DNA nanothermometer could be remotely activated and enable to perform highly sensitive and practical ratiometric temperature sensing. Meanwhile, the light-controlled DNA nanothermometer could conduct temperature sensing in the extracellular microscope and demonstrates desirable sensitivity, excellent reversibility, and quantitative ability for extracellular temperature measurement. Therefore, this light-controlled DNA can serve as a promising tool for elucidating thermal-related cell physiological and pathological processes.
Zhiwei Deng, Jiacheng Li, Hui Liu, Tong Luo, Yanjing Yang, Minghui Yang, Xiang Chen

2738 related Products with: A light-controlled DNA nanothermometer for temperature sensing in the cellular membrane microenvironment.