You identified a new mutant called wox5-2. The mutant wox5-2 protein possesses all of the activities of the wild-type WOX5 protein, except that the mutant wox5-2 protein cannot be transported into the nucleus. Answer the following questions, and briefly explain your answers. Question 6.1 Q6.1 15000 Points Grading comment: Does a homozygous wox5-2 mutant have a functional quiescent center in the root? Briefly explain your answer.
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Step 1: Identify WOX5 normal function — WOX5 is a nuclear-localized homeodomain transcription factor expressed in the quiescent center (QC) that maintains QC identity and prevents differentiation of adjacent stem cells by regulating target gene expression. Show more…
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(20 points total) You are investigating how the transcription factor BLUE1 enters into the nucleus to turn on gene transcription and expression. You introduce a green fluorescent protein tagged version of BLUE1 (called BLUE1-GFP) into wildtype cells (wt) and mutant cells with deletions in two importin proteins (ΔWELL̑α and ΔWELLβ). In each strain, you examine localization of the BLUE1-GFP tagged protein and compare it to the localization of a fluorescent molecule (DAPI) that binds to DNA. Fluorescence is indicated by the shaded regions in the diagram. a. (4 points) Examine the illustration to the right. Given these data, is either WELLα or WELLβ important for import of BLUE1 into the nucleus? Explain how you know in 2 sentences or less. (4 points) You next want to test BLUE1-GFP localization in the absence of the Ran GTPase, but you are unable to create a Ran deletion mutant. Propose an explanation for why WELLα or WELLβ deletion is able to survive, but a Ran deletion is lethal.
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A gene regulatory protein, A, contains a typical nuclear localization signal but surprisingly is usually found in the cytosol. When the cell is exposed to hormone, protein A moves from the cytosol into the nucleus, where it turns on genes involved in cell division. When you purify protein A from cells that have been treated with hormone, you find that protein B is always complexed with it. You also found that hormone treatment phosphorylates protein B. To determine the function of protein B, you express not the normal protein B but instead express a mutant form of protein B that cannot be phosphorylated. You compare normal and the cells expressing the mutant protein B using differential centrifugation to separate the nuclear fraction from the cytoplasmic fraction, and then separating the proteins in these fractions by gel electrophoresis. You identify the presence of protein A, protein B, phosphorylated form of protein B (p-protein B) and mutant protein B by looking for their characteristic bands on the gel. The gel you run is shown in the figure below
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Which mutant has a greatly increased number of stem cells in the shoot apical meristem? Group of answer choices A mutant that has NO functional CLAVATA3. A mutant that has NO functional PIN protein. A mutant that has NO functional WUS. A mutant that has NO functional WOX5.
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