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- A cloned gene fragment contains a regulatory element that isrecognized by a regulatory transcription factor. Previousexperiments have shown that the presence of a hormone resultsin transcriptional activation by this transcription factor. To studythis effect, you conduct an electrophoretic mobility shift assayand obtain the following results: Explain the action of the hormone.A. Based on this data, draw a conclusion about the promoter elements that Caudal specifically upregulates. B. Dr. Juven-Gershon also tested Caudal activation of core promoter sequences from twoother genes. Similar to the first experiment, one was DPE-dependent (from the E74Bgene), and the other was TATA-dependent (from the Adh gene). However, these two corepromoters lacked the BREU motif. The results from these additional core promoters areshown below. Based on these results and those shown above, propose a hypothesis thatexplains the differences in Caudal-mediated upregulation among all the different corepromoters Dr. Juven-Gershon tested. C. To test your hypothesis from Question 3B, you plan perform the same method of genereporter assays that Dr. Juven-Gershon has done. However, you will need to do somemutations to alter the core promoters. Make diagrams or clearly describe all the variouscore promoters mutants you wish to test. These include your controls, as well as any…Please describe the regulatory role of enhancers that binds to responsible regulators to activate the target gene at a given time and place through comparing different organisms according to the presence of regulatory sequences. regulatory promoter sequence bacteria FIGURE 19-1 The regulatory ele- ments of bacterial, yeast, and human genes. Illustrated is the increasing complex- ity of regulatory sequences from a simple bacterial gene controlled by a repressor to a human gene controlled by multiple activa- tors and repressors. In each case, a promoter is shown at the site where transcription is initiated. Although this is accurate for the bacterial case, in the eukaryotic examples, transcription initiates somewhat down- stream from where the transcription ma- chine binds (see Chapter 13). Some groups of regulatory binding sites in the human reg- ulatory sequences represent enhancers, as shown in one case. yeast human enhancer
- E27. A cloned gene fragment contains a regulatory element that is recog- nized by a regulatory transcription factor. Previous experiments have shown that the presence of a hormone results in transcriptional acti- vation by this transcription factor. To study this effect, you conduct a electrophoretic mobility shift assay and obtain the following results: Tube: 1 2 3 Transcription factor: Hormone: Explain the action of the hormone.Match whether the protein binds directly or indirectly (assuming it does) to DNA and whether to the enhancer or promoter. Items may be matched to multiple choices and not all choices will be used. 1. Directly to enhancer Repressors 2. Directly to promoter Activators 3. Indirectly to enhancer TBP associated factors 4. Indirectly to promoter >ennar region of gene X, which determines the length of the tail in mice, is mutated so that transcription factors bind it at a much higher affinity compared to the wild-type sequence. What is the most likely phenotypic outcome? Tail length will not change because the enhancer is a non-coding sequence Tail length will increased due to increased activity of the gene's promoter Tail length will decreased because any mutation will cause a loss-of-function of these regulatory regions Not just the tail will be enlarged because increased activity of the enhancer will impact many genes
- iestiðn Pre-initiation complex (PIC) is formed right before the initiation of genes, which of the following steps seemes to NOT help for the PIC formation? O TBP protein to recognize TATA box at promoter region O TFIIA to recognize the TAFS in TFIID O Enhancers to attract coactivators and activators O Silencers to attract repressor and corepressorsEach year in the United States, there are over 230,000 newcases of prostate cancer and almost 28,000 deaths. A 3.8-Mbregion on chromosome 8 (8q24), called a gene desert, has genes but contains enhancer sequences that potentiallyconfer significant risks for prostate cancer. One particular enhancerallele, which is known to be associated with an elevated risk forprostate cancer, physically interacts with the promoter region ofthe nearby MYC gene and facilitates its upregulation. Overexpressionof MYC, which encodes a cell-cycle regulatory protein, isobserved in multiple types of cancer (see Chapter 24). The riskallele has a frequency of 49 percent in men of European descentand 81 percent in men of African ancestry. Most of the differentialMYC activity associated with the risk allele occurs during prenataldevelopment, raising the possibility that testing for this alleleearly in life can be used to identify those in the African-Americanpopulation who are at very high risk for prostate…Plz do explain.Thanks Question:- Many types of breast cancer have chromosomal translocation mutations. What scenario best describes, what occurs during this type of mutation, causing cells to proliferate abnormally? Chromosomal translocations may place the gene downstream near the promoter region, therefore causing over-expression of the gene The gene may be placed in the transcription start site, downstream of the gene, initiating transcription by recruiting polymerase II Translocation mutations will initiate the transcription of mRNA in the cytoplasm of the cell catalyzing protein synthesis. Chromosomal translocations can sometimes place a gene under the control of a powerful enhancer, upstream.
- Wilms tumor 1, or nephroblastoma, is caused by mutations in the WT1 gene, which encodes a transcription factor. You have identified a novel variant in WT1: Arg422Pro. You have control cells and cells that have been engineered to carry the homozygous WT1 p.Arg422Pro mutation. You want to assess effects of this mutation on a variety of endpoints. For each endpoint listed below, choose the one technique is best suited to answer the question. Choose from: array CGH, qRT-PCR, qPCR, RNA-seq, FISH, in situ hybridization, western blot, immunostaining, WT1 ChIP-seq, WT1 ChIP-PCR, ATAC-seq, 3C Endpoint Technique? WT1 protein amount (quantitative) Western blot WT1 protein binding to all enhancers, genome-wide Chip-seq WT1 mRNA amount (quantitative) WT1 protein subcellular localization Quantitative assessment of all mRNAs in these cells (genome-wide) RNAseq Chromatin interactions between a specific WT1 chromatin binding site (identified above)…a. Would you expect a cell to continue or to stopdividing at a nonpermissive high temperature if itis a temperature-sensitive Ras mutant whose protein product is fixed in the GTP-bound form atnonpermissive temperature?b. What would you expect if you had a temperaturesensitive mutant in which the Ras protein staysin the GDP-bound form at high temperature?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.