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BIOL 429 DISC 7 - BIOL 429 Paper 7 Discussion Qu estions...

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BIOL 429 Paper 7 Discussion Questions RESULTS 2 1. Researchers prevent phosphorylation of serine 621 and see that Raf-1 remains inactive. In order to determine whether it’s the loss of phosphorylation or loss of 14-3-3 binding, they re-establish 14-3-3 binding. How do they get 14-3-3 to bind to Raf when serine 621 is not phosphorylated? 2. Is the phosphorylation of S621 required for Raf-1 activation? RESULTS 3 3. How do the mutations A259 and L257 affect the phosphorylation of Raf-1 on serine 259? 4. Do these two mutations allow binding to 14-3-3? RESULTS 4 5. Which is more important for the activation of Raf-1 by Ras: phosphorylation of S259 or binding of 14-3-3? RESULTS 5 6. Throughout the paper, what is “MBP phosphorylation” an indicator of? RESULTS 6 7. Which mutations prevent membrane localization of Raf-1? RESULTS 7 8. In Figure 7, how is a phosphorylated form of S259 identified? Researchers replaced the RSXpSXP 14-3-3 binding motif in CR3 with a phosphorylation-independent binding motif. They deleted amino acids 621 to 648 of Raf-1 and fused an R-18 peptide (which binds
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