“FEM1”nism controls SLBP stability during cell cycle

The Fluorescent Protein Stability Assay: An Efficient Method For Monitoring Intracellular Protein Stability

“…This stem-loop motif plays a key role in the maturation, export and translation of these important mRNAs, which lack polyA tails [19]. One reason for the interest in using this protein as a model is that its stability is tightly regulated along the cell cycle progression, the protein being degraded in the G1 and G2/M phases of the cell cycle [20,21]. After infection of HeLa cells with a vector expressing the FLAG-RFP-SLBP (FR-SLBP) hybrid proteins, puromycin-resistant clones were isolated.…”

“…Flow cytometry analyses of these cells after DAPI staining clearly showed that unsynchronized cells exhibited a normal cell distribution profile, whereas the synchronized cells were mainly in G2/M (Figure 3A). It has been previously shown that degradation of SLBP in G2/M is performed by the UPS [21]. Hence, the authors tested the effect of the MG132 proteasome inhibitor to see if the FR-SLBP fusion protein was sensitive to this drug.…”