CMMB 511 Lecture Notes - Lecture 17: Deoxyribonuclease I, Deoxyribonuclease, Chromatin

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rosefrog223

28 Dec 2019

School

University of Calgary

Department

Cellular, Molecular and Microbial Biology

Course

CMMB 511

Professor

Lashitew Gedamu

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Document Summary

If some sequences are transcribed and others are not transcribed in the same chromosome, even at the same tie: If so, are they distinguishable from nucleosomes in non-transcribed regions. Transcription involves the unwinding of dna and one would suspect local unfolding of chromatin fibre. One should examine the nuclear components of chromatin in transcribing and non- transcribing regions. If nucleosomal organization has changed in some region of chromatin and not in others, then some portion of dna should be exposed and in more likely to be digested with nucleases (dnase 1, monoccocal nuclease) Dnase1 could be used to probe if gene is active. Therefore, active genes should be preferentially degraded by dnase 1 than inactive genes (monococcal nuclease) Studied using probe of known sequence of gene. If you know goi and sequence, you can use sequence as probe to monitor all dna fragments that come out of gene, when it is and isnt active.

Related Questions

The bacterium E. coli, which is a major model system for the study of genes and DNA, has a singular circular chromosome of about 4x106 bp. In contrast, human diploid cells have __1__ chromosomes, of __2__ different types (donât forget that the X Â¹ Y!). The total amount of DNA found in a haploid cell such as a sperm or egg is called the organismâs âC-valueâ; the human C-value is __3__.

Also in contrast to the E. coli circular chromosome is the fact that eukaryotic chromosomes are not circular, but instead are __4__. Eukaryotic chromosomes are complexed heavily with protein to form a DNA-protein complex generically called __5__. The proteins found can be considered to fall into two general classes: __6__ and __7__. The first class is made up of 5 major proteins called __8__, __9__, __10__, __11__, and __12__. On a weight to weight basis, these proteins make up about __13__% of the total mass of a chromosome. Since these proteins must associate relatively non-specifically with DNA, they are rich in the basic amino acids __13__ and __14__, which gives the proteins a net __15__ charge.

One goal of packaging the DNA into chromatin is to compact the genome so it will fit into a nucleus. The basic unit of chromatin structure is the __16__, which has __17__ base pairs of DNA wrapped around a âcore particleâ composed of __18__ copies each of the core histones. Each nucleosome is separated by a stretch of âlinkerâ DNA so that the repeating unit is approximately 200 bp. Lets do a little arithmetic: If we consider that the nucleosome is approximately __19__ nm in diameter, that there is about 200 bp in this structure, and that 200 bp of DNA would normally occupy __20__ nm if stretched out straight (recall that each base is spaced about 0.34 nm from the next), we can see that the nucleosome alone allows for a __21__-fold compaction ratio.

The nucleosome structure is further compacted by histone __22__, which is not a part of the core particle. Instead, this histone __23__ (does what?) to cause the nucleosomal 10 nm fiber to condense into a higher-order structure referred to as the __24__ nm fiber. In the nucleus, chromosomes do not just float around freely: they are tethered to a proteinaceous structure called the __25__ at specific DNA regions called âSARsâ, which stands for __26__. SARs are rich in topoisomerases, to allow the winding and unwinding of the DNA that accompanies replication and transcription to occur, despite this constraint.

The 30 nm fiber discussed above is the form in which transcriptionally active DNA is believed to exist in the nucleus; however, further condensation leads to a highly condensed inactive form called __27__; in this terminology, the active form is referred to as __28__. The inactive condensed form can be divided into two types: one type is essentially invariant between cell types, and since it is âon all t timeâ, it is referred to as __29__. The other type comes and goes, depending on the expression of the genes present in that region of the chromosome; it is referred to as __30__.

Eukaryotic chromosomes carry the genes, of course, but also have other functional regions. One such region is the place where the spindle apparatus attaches during mitosis and meiosis; this region of the chromosome is called the __31__. Another important part of the linear eukaryotic chromosome is the ends; these are called __32__. In humans, the end of each chromosome is composed of many repeats of the short sequence __33__. One more type of functional region on each chromosome is the place where DNA replication starts; in humans, each chromosome has several of these __34__. Thus, eukaryotic chromosomes carry several different types of functional sequences, which together allow for their __35__ during S phase, __36__ during mitosis and meiosis, and the __37__ of their genes

Chapter 10

1.Outline the history of our knowledge on DNA up to Watson and Crick. What were the main contributions made by each researcherâs key experiment?

2.Explain the setup of the Hershey and Chase experiment, what would the results have been if protein was the genetic material?

3.Draw the structure of a DNA nucleotide, labeling each main component correctly. How does an RNA nucleotide differ?

4.If a section of double stranded DNA contains 19% Adenine, how much Thymine is present?

5.You are a researcher studying the genetic basis of heart attacks and have been working to determine the expression levels of different genes that might contribute to cancer formation. You obtain the DNA methylation status of five genes of interest (the data are shown in the table below). The plus (+) sign indicates the level of DNA methylation; more plus signs correlates with increased methylation levels.Based on this information which genes would you predict to have the highest rate of transcription?

Gene	Methylation levels
1	++
2	+++++
3	+++
4	++
5	+

What are the characteristics of the 3 main DNA forms?

Chapter 11

What are the different types of chromatin?

What are the structures and important roles for telomeres and centromeres?

What are the differences found between eukaryotic chromosomes and mitochondrial?

Chapter 12

Explain each of the different models of replication.

If you grow a culture of bacteria in media with radioactive nucleotides so that all DNA in the cells include radioactive nucleotides and then place the bacteria in new non radioactive media. After two rounds of replication what proportion of the DNA molecules will contain radioactivity?

Summarize the similarities and differences between rolling-circle replication, theta replication and linear eukaryotic replication.

What are the functions of the different DNA polymerases found in eukaryotic cells?

Draw a replication fork and include all key components and orientations. (Leading/lagging strands, DNA helicase, RNA primer and DNA gyrase)

What is the Holliday model of recombination and what are the necessary steps?

Chapter 13

What are the different types of RNA and what roles do they play?

Describe the properties and functions of each of the RNA polymerases and how they differ depending on the organism.

Describe in detail the process and mechanisms of transcription in both prokaryotes and eukaryotes.

Chapter 14

What are the primary purposes of each of the three post transcriptional modifications that occur in eukaryotic cells.

What is alternative splicing and what role does it play in the cell?

How is ribosomal RNA processed after transcription?

How do siRNA and miRNA work, describe/draw out the process in detail.

QUESTION 10

If you can drink milk as an adult, it means that you have inherited a mutation in the promoter of your lactase gene (the gene that encodes the enzyme you need to break down lactose). Predict the effect of this mutation:

		The mutation changes the number of domains in the enzyme, which makes it work more efficiently
		The mutation changes the amino acid sequence of the lactase protein
		The mutation increases the number of copies of the lactase gene that will be found in your genome
		The mutation changes whether the lactase sequence is found in an intron or exon
		The mutation affects the expression of the lactase gene

1.2 points

QUESTION 11

A competitive inhibitor is decreasing the activity of an enzyme. Predict the effect of adding more substrate to the reaction.

		The substrate will increase the reaction rate by binding to the allosteric site
		The substrate will increase the reaction rate by competing with the inhibitor for the active site
		The reaction rate will not change unless the inhibitor can be removed
		The enzyme adjusts its shape so that the substrate, but NOT the competitive inhibitor, can bind
		The substrate will bind to the competitive inhibitor and block its ability to bind to the enzyme

1.2 points

QUESTION 12

What determines where in the genome a transcription regulator will bind?

		Transcription regulators bind to the 5' UTR region of a gene
		Regulators bind via complementary base-pairing to certain DNA molecules
		Covalent bonds form between the transcription regulator and the atoms of the DNA backbone
		Every eukaryotic gene has a different transcription regulator that will bind to the 5' end of the gene
		Transcription regulators bind to specific DNA sequences via multiple weak non-covalent interactions

1.2 points

QUESTION 13

What is the basic premise of cell theory?

		DNA -> RNA -> protein
		All cells arise from pre-existing cells
		DNA provides the complete instructions to create a cell
		The identity of a cell is determined through gene expression patterns
		All cells contain the same four basic macromolecules

1.2 points

QUESTION 14

What is the benefit of using BOTH the lac activator and the lac repressor to control gene expression?

		Using both an activator and repressor enables cells to more accurately determine the amount of lactose available in the environment
		Enzymes to digest lactose are only made when energy is low and lactose is available
		The activator can override the inhibition of the lac operon by the repressor
		The repressor can control the enhancer, while the activator can control the promoter
		When neither the lac activator or repressor is present, expression of the lac operon is too high

1.2 points

QUESTION 15

What is the histone code used for?

		Phosphorylation and acetylation of DNA affect its ability to be compacted
		Changes to the sequence of DNA change whether DNA will wrap around histone proteins
		Covalent modifications of histones affect the ability of the transcription initiation complex to form
		Histones provide the codon sequences needed for translation to occur
		The histone code affects which amino acids will get added to a polypeptide

CMMB 511 Lecture Notes - Lecture 17: Deoxyribonuclease I, Deoxyribonuclease, Chromatin

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