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- The retinoblastoma tumor suppressor gene Rb (RB1) codes forthe retinoblastoma protein (pRB). pRB prevents the progression of the cell cycle through G1 if DNA has been damaged. Itdoes so in part because it binds a transcription-activatingdimer referred to as E2F-DP. The pRB-E2F/DP complex recruits a histone deacetylase to chromatin. Explain.In tumor cells obtained from patients with Burkittlymphoma, a cancer of the immune system’s B cells,the myc gene often appears close to one of the breakpoints of a reciprocal translocation between chromosomes 8 and 14. In this translocated position, myc is expressed at a higher-than-normal level. Scientists hypothesize that Myc protein overexpression in B cellscontributes to lymphoma formation.a. Explain how transgenic mice produced using pronuclear injection could be used to test this hypothesis. (Assume that you previously cloned a generegulatory region that is active specifically in Bcells throughout the life of the mouse.)b. Suppose you wanted to overexpress Myc only inthe immune cells of mice, starting at one week ofage. To restrict Myc transcription spatially, youwill use same promoter described in part (a). Torestrict Myc transcription temporally, you will usea cre transgene whose expression is controlled byheat shock (hs-cre). Describe the mouse you wouldcreate to accomplish…Recall that the nuclear membrane disintegrates late in prophase of mitosis in most eukarvotic cells. Once the membrane is reformed in telophase in a daughter cell, several components of gene expression mignt therefore be "caught" out in the cytoplasm when they would otherwise onlv ever be found inside the nucleus. Consider where the following components of gene expression are made and where they runction. Which component is normally never found in the cytoplasm outside the nucleus? O A. Spliced intron • B. RNA polymerase O C. Histones • D. DNA polymerase
- Cell lines divide normally in a defined medium containing growth factors, but fail to divide in the absence AGF (a growth factor). However, a mutant cell line continues to divide even in the absence of AGF. Elevated levels of Rb phosphorylation and the effects of receptor and Mek inhibitors suggest a mutation activating an oncogene. Inhibitors of Mek inhibit cell division of the mutant cell line, but inhibitors to the ADGF receptor, a receptor tyrosine kinase (RTK) with homology to EGFR, do not. Outline the RTK pathway leading to the phosphorylation of Rb to form p-Rb.Regulation of the cell cycle is very complex and involves multiple proteins. In yeast, a complex of cdc2 and a mitotic cyclin is responsible for moving the cell past the G2/M checkpoint. The activity of the cyclin-dependent kinase cdc2 is inhibited when it is phosphorylated by the kinase, Wee-1. What would you predict would be the phenotype of a Wee-1 mutant yeast? What other genes could be altered in a Wee-1 deficient mutant strain that would make the cells act normally?Describe the effects of the mutation causing the p21 promoter to no longer bind p53 on cell signaling pathways and metabolism or cell cycle control.
- You are studying Protein X which plays a role in promoting the G1/S phase transition in eukaryotic cells. You design an experiment using wild-type yeast cells to measure the amount of Gene X MRNA and activity levels of Protein X during the cell cycle. The results from your experiment are shown in the graph below. From the data, which of the following would most likely result from increased activity of Protein X? Select all that apply - Protein X activity levels Gene X MRNA levels Relative Units G1 G2 M Activation of the anaphase promoting complex (APC/C) and cleavage of cohesin proteins Activation of proteins that bind to Ori (origin of replication) sites Increased levels of dynamic instability of microtubules O Increased phosphorylation of nuclear lamin proteins, leading to breakdown of nuclear lamina structureThe gene that encodes the enzyme called tyrosine hydroxylase is known to be activated by the CREB protein. Tyrosine hydroxylase is expressed in nerve cells and is involved in the synthesis of catecholamine, a neurotransmitter. The exposure of cells to adrenaline normally up-regulates the transcription of the tyrosine hydroxylase gene. A mutant cell was identified in which the tyrosine hydroxylase gene was not up-regulated when exposed to adrenaline. List all the possible mutations that could explain this defect. How would you explain the defect if only the tyrosine hydroxylase gene was not up-regulated by the CREB protein, whereas other genes having CREs were properly up-regulated in response to adrenaline in the mutant cell?In addition, when ARF is artificially expressed in normal cells, a rapid increase in p53 levels is observed. Studies have demonstrated that ARF, when expressed, is associated with Mdm2. When associated, the two proteins display a particular pattern of localization restricted to the nucleolus. Suggest a mechanism by which ARF leads to p53 build-up.
- Explain the role of CDK inhibitors. If cyclin-CDK complexes are necessary to allow regulated progression through the eukaryotic cell cycle, what would be the physiological rationale for CDK inhibitors?The PYK gene codes for the expression of pyruvate kinase, which is one of the enzymestargeted for anti-cancer drug design. You have identified an RNAi that targets the mRNAof PYK gene. To study the effect of the RNAi towards pyruvate kinase, the respected RNAiis expressed in Saccharomyces cerevisiae. The level of pyruvate kinase can be detectedwith a fluorescent antibody.(a). Predict the result that you will obtain in recombinant S. cerevisiae that expresses therespected RNAi.(b). Compare the result in Q3a(i) with the wild-type S. cerevisiae.3) The tumor suppressor protein Rb regulation of the entry into the S phase of the cell cycle is represented in this diagram. DNA Answer: b) Explain your choice above: Answer: Rb E2F Genes needed for S phase are NOT transcribed Growth factor Ras pathway Cdk-cyclin 30 ATP ADP Phosphorylated Rb protein P Rb E2F Gene transcription a) In hereditary retinoblastoma tumors, Rb is mutated. Among the following mutations, which one is not likely to be found in these tumors. 1) Mutation prevents Rb to bind E2F by modifying the binding site. 2) Mutation prevents Rb to be dephosphorylated and recycled (possibly by prevented phosphorylated Rb to be recognized by the phosphatase that removes its phosphates). 3) Mutation may cause Rb to be misfolded and not have a functional conformation 4) Mutation that prevent Rb to be phosphorylated by cdk-cyclin. 5) Mutation may cause Rb to be unstable and degraded rapidly. c) (4 pts) Human papilloma virus (HPV) infections are the main causes of cervical cancers.…