Figure 5.

Loss of Tsc1 results in greater polysomes and increased rates of protein synthesis. A, 3 × 10⁶ cells for each genotype were lysed and fractionated over a 10% to 45% sucrose gradient. Gradients were fractionated and ribosomal subunits were detected by measuring RNA absorbance at 254 nm. Percentages are given for differences in calculated areas under each polysome peak. B, [³⁵S]methionine was added to methionine-starved cells of each genotype for the indicated times. Cells were immediately lysed and total protein was precipitated with TCA, pelleted, and subjected to liquid scintillation counting to measure incorporated cpm.