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Ch. 17+18 - Transcriptional Regulation in Eukaryotes
Chapter 17, Problem 22

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A deletion within the GAL4 gene that removes the region encoding amino acids 1 to 100.

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1
Understand the role of the GAL4 gene: GAL4 is a transcriptional activator in yeast that binds to the upstream activating sequence (UAS) of the GAL1 gene, facilitating transcription in the presence of galactose.
Identify the significance of amino acids 1 to 100 in GAL4: This region includes the DNA-binding domain, which is crucial for GAL4 to attach to the UAS of the GAL1 gene.
Consider the effect of the deletion: Removing amino acids 1 to 100 would likely disrupt the DNA-binding ability of GAL4, preventing it from binding to the UAS.
Predict the impact on transcription: Without GAL4 binding to the UAS, the transcription of the GAL1 gene would be significantly reduced or completely inhibited, even in the presence of galactose.
Conclude the overall effect: The deletion within the GAL4 gene would impair the activation of GAL1 transcription, highlighting the importance of the DNA-binding domain in gene regulation.

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Key Concepts

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

GAL4 Protein Function

GAL4 is a transcription factor in yeast that binds to specific DNA sequences in the promoter region of the GAL1 gene. It activates transcription in the presence of galactose by recruiting the transcriptional machinery. A deletion in the GAL4 gene that removes the first 100 amino acids would likely disrupt its ability to bind DNA or interact with other proteins, leading to reduced transcription of the GAL1 gene.
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Transcription Regulation

Transcription regulation involves mechanisms that control the rate of gene expression. In yeast, the presence of galactose activates the GAL pathway, where GAL4 plays a crucial role. Mutations that affect transcription factors like GAL4 can significantly alter the expression levels of target genes, such as GAL1, impacting the organism's ability to metabolize galactose.
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Mutations and Their Effects

Mutations are changes in the DNA sequence that can affect gene function. In this case, a deletion mutation in the GAL4 gene can lead to a loss of function, which may prevent the proper activation of the GAL1 gene. Understanding how specific mutations impact protein structure and function is essential for predicting their effects on transcription and overall cellular metabolism.
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Related Practice
Textbook Question

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A mutation within the GAL80 gene that blocks the ability of Gal80 protein to interact with Gal3p.

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Textbook Question

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A deletion of one of the four UASG elements upstream from the GAL1 gene.

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Textbook Question

What role do ubiquitin ligases play in the regulation of gene expression?

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Textbook Question

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A deletion of the entire GAL3 gene.

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Textbook Question

Much of what we know about gene interactions in development has been learned using nematodes, yeast, flies, and bacteria. This is due, in part, to the relative ease of genetic manipulation of these well-characterized genomes. However, of great interest are gene interactions involving complex diseases in humans. Wang and White [(2011). Nature Methods 8(4):341–346] describe work using RNAi to examine the interactive proteome in mammalian cells. They mention that knockdown inefficiencies and off-target effects of introduced RNAi species are areas that need particular improvement if the methodology is to be fruitful.

How might one use RNAi to study developmental pathways?

257
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Textbook Question

Much of what we know about gene interactions in development has been learned using nematodes, yeast, flies, and bacteria. This is due, in part, to the relative ease of genetic manipulation of these well-characterized genomes. However, of great interest are gene interactions involving complex diseases in humans. Wang and White [(2011). Nature Methods 8(4):341–346] describe work using RNAi to examine the interactive proteome in mammalian cells. They mention that knockdown inefficiencies and off-target effects of introduced RNAi species are areas that need particular improvement if the methodology is to be fruitful.

Comment on how 'knockdown inefficiencies' and 'off-target effects' would influence the interpretation of results.

354
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