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Ch. 10 - Eukaryotic Chromosome Abnormalities and Molecular Organization
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 10, Problem 25

Experimental evidence demonstrates that the nucleosomes present in a cell after the completion of S phase are composed of some 'old' histone dimers and some newly synthesized histone dimers. Describe the general design for an experiment that uses a protein label such as ³⁵S to show that nucleosomes are often a mixture of old and new histone dimers following DNA replication.

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Design an experiment where cells are grown in a medium containing ³⁵S-labeled methionine during the S phase of the cell cycle. This will incorporate the radioactive label into newly synthesized histone proteins.
After the S phase, isolate the chromatin from the cells to analyze the nucleosomes.
Use a method such as SDS-PAGE followed by autoradiography to separate and visualize the histone proteins. This will allow you to detect the presence of ³⁵S-labeled histones.
Compare the labeled histones to unlabeled histones by using a control group of cells grown without ³⁵S-labeled methionine. This will help determine the proportion of new histones incorporated into nucleosomes.
Analyze the results to determine the presence of both labeled (new) and unlabeled (old) histone dimers in the nucleosomes, indicating a mixture of old and new histones after DNA replication.

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Key Concepts

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

Nucleosome Structure

Nucleosomes are the fundamental units of chromatin, consisting of DNA wrapped around a core of histone proteins. Each nucleosome is made up of two copies of four different histone proteins (H2A, H2B, H3, and H4), forming an octamer. Understanding nucleosome structure is essential for grasping how histones can be old or newly synthesized after DNA replication.
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Chromosome Structure

DNA Replication and Histone Dynamics

During DNA replication, the original histones are distributed between the two daughter strands, while new histones are synthesized to fill in the gaps. This process results in nucleosomes that contain a mix of old and new histone dimers. Recognizing this dynamic is crucial for designing experiments that track histone incorporation into nucleosomes post-replication.
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Protein Labeling Techniques

Protein labeling techniques, such as using radioactive isotopes like ³⁵S, allow researchers to trace the incorporation of specific proteins into cellular structures. By labeling newly synthesized histones, one can distinguish between old and new histone dimers in nucleosomes. This method is vital for demonstrating the mixture of histone types in nucleosomes after DNA replication.
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Related Practice
Textbook Question

A boy with Down syndrome (trisomy 21) has 46 chromosomes. His parents and his two older sisters have a normal phenotype, but each has 45 chromosomes.

How many chromosomes do you expect to see in karyotypes of the parents?

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Textbook Question

A boy with Down syndrome (trisomy 21) has 46 chromosomes. His parents and his two older sisters have a normal phenotype, but each has 45 chromosomes.

What term best describes this kind of chromosome abnormality?

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Textbook Question

A boy with Down syndrome (trisomy 21) has 46 chromosomes. His parents and his two older sisters have a normal phenotype, but each has 45 chromosomes.

What is the probability the next child of this couple will have a normal phenotype and have 46 chromosomes? Explain your answer.

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Textbook Question

DNase I cuts DNA that is not protected by bound proteins but is unable to cut DNA that is complexed with proteins. Human DNA is isolated, stripped of its nonhistone proteins, and mixed with DNase I. Samples are removed after 30 minutes, 1 hour, and 4 hours and run separately in gel electrophoresis. The resulting gel is stained to make all DNA fragments in it visible, and the results are shown in the figure. DNA fragment sizes in base pairs (bp) are estimated by the scale to the left of the gel. Examine the gel results and speculate why longer DNase I treatment produces different results.

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Textbook Question

DNase I cuts DNA that is not protected by bound proteins but is unable to cut DNA that is complexed with proteins. Human DNA is isolated, stripped of its nonhistone proteins, and mixed with DNase I. Samples are removed after 30 minutes, 1 hour, and 4 hours and run separately in gel electrophoresis. The resulting gel is stained to make all DNA fragments in it visible, and the results are shown in the figure. DNA fragment sizes in base pairs (bp) are estimated by the scale to the left of the gel. Draw a conclusion about the organization of chromatin in the human genome from this gel.

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Textbook Question

Genomic DNA from the nematode worm Caenorhabditis elegans is organized by nucleosomes in the manner typical of eukaryotic genomes, with 145 bp encircling each nucleosome and approximately 55 bp in linker DNA. When C. elegans chromatin is carefully isolated, stripped of nonhistone proteins, and placed in an appropriate buffer, the chromatin decondenses to the 10-nm fiber structure. Suppose researchers mix a sample of 10-nm–fiber chromatin with a large amount of the enzyme DNase I that randomly cleaves DNA in regions not protected by bound protein. Next, they remove the nucleosomes, separate the DNA fragments by gel electrophoresis, and stain all the DNA fragments in the gel.

Approximately what range of DNA fragment sizes do you expect to see in the stained electrophoresis gel? How many bands will be visible on the gel?

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