3. In addition to cyclin binding, there is a second mechanism that keeps the G1 CDK inactive. What is it? Why do you think this second layer of regulation is needed?
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- 2. The mannose 6-phosphate receptor is located predominantly in endosomes and the TGN, but about 10% of the mannose 6-phosphate receptor molecules are in the plasma membrane. Suppose you have an antibody that associates with the mannose 6-phosphate receptor and blocks the ability of the receptor to bind mannose 6-phosphate. Suppose further that this association is rapid and effectively irreversible. If you incubate cells with the antibody for a brief time and then wash away excess antibody, the cells continue to sort lysosomal enzymes normally. But if you incubate cells continuously with the antibody for a prolonged time, the cells begin to secrete lysosomal enzymes. a) How would you interpret this result? b) Suppose that the antibody has been fluorescently labeled by covalently attaching a fluorophore. If you examine cells that have been incubated continuously with this labeled antibody, what types of fluorescence patterns would you expect to see at different time points after…See figure 12.16b regarding the process by which cyclin regulates the Cdk. Suppose that the cyclin binding site in the Cdk contains these FOUR amino acids in this order from top to bottom: serine, lysine, aspartic acid acid and lysine and the Cdk binding site in the cyclin contains these FOUR amino acids in this order from top to bottom: aspartic acid, aspartic acid, lysine and serine. Use the schematics below to show the R groups and how they might interact to create the cyclin.cdk complex. Label both binding sites, show all charges that will be used to create any bonds, and label all bonds formed and add the ATP active site. Cyclin Serine Lysine Aspartic "Acid Lysine -OH NH3t -Coo NH3+ NH3 + -OH Aspartic Aad Aspartic Acid /Lysine Serine сокSee figure 12.16b regarding the process by which cyclin regulates the Cdk. Suppose that the cyclin binding site in the Cdk contains these FOUR amino acids in this order from top to bottom: serine, lysine, aspartic acid acid and lysine and the Cdk binding site in the cyclin contains these FOUR amino acids in this order from top to bottom: aspartic acid, aspartic acid, lysine and serine. Use the schematics below to show the R groups and how they might interact to create the cyclin.cdk complex. Label both binding sites, show all charges that will be used to create any bonds, and label all bonds formed and add the ATP active site. A Explain what a kinase does and how the cyclin controls the activity of the Cdk.
- Figure 9.10 In certain cancers, the GTPase activity of the RAS G-protein is inhibited. This means that the RAS protein can no longer hydrolyze GTP into GDP What effect would this have on downstream cellular events?3. The figure to the right shows the changes in the level of a cyclin and activity of cyclin-Cdk during the cell cycle. Note how the level of cyclin increases slowly & steadily, whereas the activity of the cyclin-Cdk appears quite suddenly. How would we best explain the rapid activation of cyclin-Cdk? a. Cyclin-Cdk gene expression leads to rapid protein synthesis. b. All cyclin-Cdk is activated when it exits the nucleus. c. Ubiquination leads to proteolytic activation of the cyclin. d. Positive feedback loops cause its rapid activation. Cyclin-Cdk Cyclin G1 S G2 M# 3 You've engineered a mutant cell where the FADD adapter was truncated. The mutant FADD only contains the Death Domain, and lacks the Death Effector Domain. What is the most likely phenotypic outcome for this mutant cell when presented with the Fas ligand? 20 E O The Fas/FasL oligomer is formed, but apoptosis is blocked O The Fas/FasL oligomer is formed, and apoptosis is hyperactivated O The Fas/FasL oligomer is not formed, and apoptosis is blocked O The Fas/FasL oligomer is not formed, but apoptosis is hyperactivated F3 $ 4 DOD 000 R F4 % 5 F5 T BARAT 6 tv @ MacBook Air F6 Y & 7 F7 U * ➤11 8 F8 · 9 F9
- 1. Which of the following signaling events is NOT engaged downstream of phospholipase C (PLC)-gamma activation? a. Calcium ion (Ca2+) release from the endoplasmic reticulum (ER) b. Heterodimerization of the subunits c-Fos and c-Jun to form the transcription factor AP-1 c. IkappaB degradation and resulting translocation of NFkappaB into the nucleus d. Recruitment of protein kinase C (PKC)-theta to the plasma membrane e. PIP3 generation at the plasma membrane which recruits and activates Akt, leading to cytoskeletal remodeling1. The following image shows a mechanism in which gene expression activity is regulated by ligand. Arg RS-2 Teu Met RBS Ligand RBS hidden a. What is this kind of regulatory machnism called? b. Does it involve transcription or translation? c. What happens in the presence of the ligand? d. What happens in the absence of theligand? e. What do you think the genes that are regulated here - metabolic (breakdown) or anabolic (buildup) for the ligand? Explain1A.If transcription factor G is downstream of EGFR and upregulates gene H, what happens to H if Grb2 has an ER signal sequence? 1B. If transcription factor A is phosphorylated by Akt and upregulates gene B, what happens to gene B if a PI3K inhibitor is added? Choices are: A. No expression of the gene B. Upregulation even in the absence of stimulation/ EGF C. Upregulation but only in the presence of cytokine/ EGF
- 14)How does a compound that inhibits the GTPase activity of ras affect cell responses to growth factors? It would increase proliferation. It would increase glucose production. It would decrease glucose production. It would decrease proliferation.Explain how CDK activity is modulated by the following proteins: (a) cyclin, (b) CAK, (c) Wee1, (d) p21.8. Describe the MAP kinase activation pathway
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