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Ch. 16+17 - Transcription, RNA Processing, and Translation
Chapter 16, Problem 16

The primary cause of death from αα-amanitin poisoning is liver failure. Suppose a physician informs you that liver cells die because their rate of protein production falls below a level needed to maintain active metabolism. Given that αα-amanitin is an inhibitor of transcription, you wonder if this information is correct. Propose an experiment to determine whether the toxin also has an effect on protein synthesis.

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Design an experiment with two groups of liver cells: one treated with αα-amanitin and one control group without the toxin. This will help determine if the toxin affects protein synthesis directly beyond its known effect on transcription.
Measure the baseline protein synthesis levels in both groups before introducing αα-amanitin to ensure that both groups start with similar protein production rates.
Expose the experimental group of liver cells to αα-amanitin while keeping the control group toxin-free. Maintain both groups under identical conditions to ensure that any differences observed are due to the toxin.
At various time intervals after exposure, measure the rate of protein synthesis in both groups. This can be done using radiolabeled amino acids that are incorporated into newly synthesized proteins, which can then be quantified.
Compare the protein synthesis rates between the toxin-treated and control groups. A significant reduction in protein synthesis in the αα-amanitin treated group, compared to the control group, would suggest that the toxin has a direct effect on protein synthesis, in addition to its inhibition of transcription.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Transcription and Translation

Transcription is the process by which messenger RNA (mRNA) is synthesized from a DNA template, while translation is the subsequent process where ribosomes synthesize proteins based on the sequence of the mRNA. Understanding these processes is crucial because αα-amanitin inhibits RNA polymerase II, which is responsible for mRNA synthesis, thereby directly affecting protein production.
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Review of Transcription vs. Translation

Protein Synthesis

Protein synthesis involves the creation of proteins through the processes of transcription and translation. It is essential for cell function and survival, as proteins perform a variety of roles, including enzymatic activity and structural support. A decrease in protein synthesis can lead to cell dysfunction and death, particularly in metabolically active cells like liver cells.
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Experimental Design

Experimental design refers to the planning of an experiment to test a hypothesis effectively. In this context, it involves setting up a controlled experiment to measure the effects of αα-amanitin on protein synthesis, possibly by using techniques such as Western blotting or measuring mRNA levels, to determine if the toxin impacts the translation process or the availability of mRNA.
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