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PCR Cloning and Subcloning> | … Anti-S6K (GenScript), anti-4E-BP1 T70, anti-4E-BP1 S65 (Abcam) and anti-Raptor were from Cell Signaling Technologies, anti-HA (Ascites HA-7 Clone … of cells, 1 × 10 6 cells were plated onto a 60-mm dish 24 h prior to transfection of 4 μg of DNA using Lipofectamine reagent … | Get A Quote |
The cap dependent translation initiation is a tightly controlled process of cooperative ternary complex formation by 4E-BP1, eIF4E and the 5' cap of eukaryotic mRNA in response to environmental cues like glucose, nutrients and growth factor levels. Based on the well-described effects of mTORC1/rapamycin complex on 4E-BP1 phosphorylation/s, it is generally accepted that rapamycin is a global inhibitor of cap-dependent translation. We have previously shown that 4E-BP1 resistance to rapamycin was overcome by the stoichiometric abundance of S6K1. Now we present evidence that the TOS-bearing amino terminal domain of S6K1 is sufficient to relieve the rapamycin resistance of 4E-BP1 as TOS deleted variants of S6K... More