your combined answers to all of these questions should be

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Read the short review article by Divakaruni and Murphy, as well as the primary research article: “A Mitochondrial Pyruvate Carrier Required for Pyruvate Uptake in Yeast, Drosophila, and Humans,” by Bricker, D.K., et al., (2012)

You must only include your answers, do not repeat the questions. Moreover, DO NOT a)copy (or quote) from the journal article, b) copy from any internet source that you find or subscribe to; all of these activities are clearly plagiarism. You need to use your own words for all answers.

1. Bricker et al., provide data from a number of experiments supporting their claim that MPC1, MPC2, and MPC3 are components of the yeast pyruvate transporter. Describe three distinct experimental approaches presented in Figure 1 indicating MPC1 is a major component of a highly conserved mitochondrial pyruvate transporter complex, whereas MPC3 provides redundant function. Note that five different types of experiments were used and you just need to describe three.

2. The authors have studied several families that display defects in mitochondrial pyruvate oxidation, which led them to test their hypothesis that one or more of these families may in fact have a genetic defect in t he human MPC1 pyruvate transporter. The data from these studies are shown in Figure 4.

2a. Family #1 showed a severe defect in mitochondrial pyruvate oxidation, whereas families #2 and #3 had less severe phenotypes. How was the genetic defect discovered in these families, and what did DNA sequencing reveal that led the authors to propose that the disease phenotype was due to MPC1 mutations?

2b. Isolated fibroblasts from these patients were tested in pyruvate-driven respiration assays in which O2 consumption was used as a measure of mitochondrial activity. What was observed when fibroblasts from families #1 and #2 were compared to fibroblasts from normal individuals with respect to basal O2 consumption in the presence of pyruvate and glutamine as compared to O2 consumption in the presence of these same metabolites and FCCP? Explain.

 

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