MBG 2040 Lecture Notes - Lecture 6: Tobacco Mosaic Virus, Ribonuclease, Deoxyribonuclease

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rubycheetah358

2 May 2018

School

University of Guelph

Department

Molecular Biology and Genetics

Course

MBG 2040

Professor

Krassimir Joseph Yankulov

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Structure of DNA and Chromosomes

Outline:

• Functions of genetic material

• Structures of DNA and RNA

• Chromosome structure in prokaryotes and viruses

• Chromosome structure in eukaryotes

Outcomes:

• Explain how genetic information is stored in deoxyribonucleic acid (DNA)

• Define the composition and structure of DNA

• Explain how DNA is organized and packaged in prokaryotes and eukaryotes

The genetic material must replicate, control the growth/development of the organism, and allow the

organism to adapt to changes in the environment.

Evidence that DNA is the genetic material:

• Recall bacterial transformation

• *see slide

• 1944: Avery, MacLeod, and McCarty determined what type of molecule is the "transforming

principle" in S. pneumoniae

o Type IIR cells --> IIR colonies

o Type IIR Cells + protein from heat-killed type IIS cells --> IIR colonies

o Type IIR Cells + DNA from heat-killed type IIS cells --> IIS colonies

o Type IIS cell DNA + protease --> IIS colonies

o Type IIS cell RNA + RNAase --> IIS colonies

o Type IIS cell DNA + DNAase --> no colonies

*Conclusion: the hereditary "transforming principle" is DNA

Evidence that RNA is genetic material:

• Tobacco mosaic virus (TMV) has RNA and protein, but no DNA

• Different strains have different coat proteins

• RNA specifies the coat protein

Key Points:

• Genetic information of most organisms is stored in deoxyribonucleic acid (DNA)

• In some viruses, the genetic information is present in ribonucleic acid (RNA)

Structure of DNA and RNA:

• Nucleic acids are composed of nucleotides

o Nitrogen containing bases:

• Pyrimidines:

▪ Thymine (in DNA)

▪ Cytosine

▪ Uracil (in RNA)

• Purines:

▪ Adenine

▪ Guanine

• Sugar (Pentose)

o Deoxyribose (-H *no hydroxyl group)

o Ribose (-OH)

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Document Summary

Outline: functions of genetic material, structures of dna and rna, chromosome structure in prokaryotes and viruses, chromosome structure in eukaryotes. Outcomes: explain how genetic information is stored in deoxyribonucleic acid (dna, define the composition and structure of dna, explain how dna is organized and packaged in prokaryotes and eukaryotes. The genetic material must replicate, control the growth/development of the organism, and allow the organism to adapt to changes in the environment. Evidence that rna is genetic material: tobacco mosaic virus (tmv) has rna and protein, but no dna, different strains have different coat proteins, rna specifies the coat protein. Key points: genetic information of most organisms is stored in deoxyribonucleic acid (dna) In some viruses, the genetic information is present in ribonucleic acid (rna) In vivo dna is negatively supercoiled: *see slide, relaxed, nicked, circular dna undergoes one 360 degree left-handed rotation --> negatively supercoiled, covalently closed dna.

Related Questions

In addition to identifying the genetic material, the experiments of Avery, MacLeod, and McCarty with different strains of Streptococcus pneumoniae demonstrated that

	DNA is a double helix.
	DNA may be taken up by bacterial cells and be active.
	DNA is present in bacteriophage T2.
	eukaryotic cells may be genetically transformed.
	DNA binds to the cell wall of the bacterium.

In order to show that DNA in cell extracts is responsible for genetic transformation in Streptococcus pneumoniae, important corroborating evidence should indicate that _______ also destroy transforming activity.

	temperatures high enough to kill cells
	enzymes that hydrolyze proteins
	enzymes that hydrolyze DNA
	enzymes that hydrolyze polysaccharides
	enzymes that hydrolyze RNA

Based on what you have learned about the experiments conducted by Griffith and Avery and colleagues with bacteria, which of the following would result in transformation of living R cells?

	Pure RNA from S cells
	Cell-membrane extracts from S cells
	A mixture of pure RNA and protein from S cells
	Cell-free extracts from S cells
	Pure protein from S cells

A-T base pairs in a DNA double helix

	form three hydrogen bonds with each other.
	are more heat-stable than G-C base pairs.
	are of a substantially different length than G-C base pairs.
	are not accessible to DNA binding proteins.
	are chemically distinct from G-C base pairs.

If 23 percent of the bases in a sample of double-stranded DNA are adenine, what percentage of the bases are uracil?

	27
	50
	0
	73
	23

The uniform diameter of the DNA structure provides evidence for

	antiparallel DNA strands.
	a right-handed helix.
	a double-stranded helix.
	3â² end phosphorylation.
	purineâpyrimidine pairing.

If a sequence of one strand of DNA is 5â²-TGACTATC-3â², what is the complementary strand?

	5â²-ACTGATAG-3â²
	3â²-TGACTATC-5â²
	3â²-ACTGATAG-5â²
	3â²-ACTGATAG-3â²
	5â²-TGACTATC-3â²

What structural aspect of the DNA facilitates dissociation of the two DNA strands for replication?

	3â² end phosphorylation
	Purineâpyrimidine pairing
	Antiparallel DNA strands
	Covalent bonds between sugars and phosphate groups
	Hydrogen bonding between paired bases

If the MeselsonâStahl density gradient experiment had resulted in two bands of DNA molecules after only one round of replication, one containing only ¹⁵N and the second only ¹⁴N, this result would have indicated that replication was

	unidirectional.
	dispersive.
	conservative.
	bidirectional.
	semiconservative.

10.

The nucleoside analogue acyclovir, which is used to treat herpes simplex virus (HSV) infections, lacks a 3â² hydroxyl group (âOH). Predict what will happen if the host cell DNA polymerase incorporates a molecule of acyclovir into an elongating strand of HSV DNA.

	The replication complex will no longer bind to origins of replication.
	The DNA polymerase will only incorporate acyclovir molecules.
	Acyclovir will bind single-strand binding proteins.
	DNA polymerase will not be able to link a successive nucleotide.
	Acyclovir will block the activity of the DNA helicase.

11.

Which of the following does not demonstrate the stability of the DNA double helix?

	Single-strand binding proteins are required to keep the strands apart.
	Paired bases form hydrogen bonds.
	High temperatures are required to denature it.
	DNA synthesis requires energy.
	DNA helicases require ATP.

12.

What effect would a primase inhibitor have on DNA replication?

	DNA polymerase would not be able to add bases to the DNA.
	Primase would be blocked from joining the DNA replication complex.
	Primase would not be able to provide primers for DNA polymerases.
	DNA polymerase would not be able to use DNA as a template.
	There would be no effect on DNA replication.

13.

To replicate their DNA in a timely manner, most eukaryotic chromosomes

	normally have uncoiled DNA.
	have multiple origins of replication.
	contain linear molecules of single-stranded DNA.
	have two replication forks that move in the same direction.
	are composed entirely of DNA.

14.

Which statement about DNA replication is false?

	Okazaki fragments are synthesized as parts of the leading strand.
	Error rates for DNA replication are reduced by proofreading of the DNA polymerase.
	The sliding clamp increases the rate of DNA synthesis.
	Replication forks represent areas of active DNA synthesis on the chromosomes.
	Ligases and polymerases function in the vicinity of replication forks.

15.

In many eukaryotes, there are repetitive sequences called telomeres at the ends of chromosomes. After successive rounds of DNA replication, the _______ strand becomes shorter. In some cells, an enzyme called _______ repairs the shortened strand.

	leading; telomerase
	lagging; telomerase
	lagging; DNA polymerase I
	leading; DNA polymerase I
	leading; replicase

16.

A researcher studies normal human fibroblast cells. They can be maintained in culture but die off after about 30 cell generations. Unexpectedly, a colony of cells continues to survive and divide past 30 generations. Which scenario is most likely true for these cells?

	DNA polymerase is constantly active.
	Primase is able to extend the telomeres.
	The mismatch repair system is extra efficient.
	The telomerase gene is being expressed.
	The cells do not need the ends of their chromosomes.

17.

If DNA polymerase III introduces an incorrect nucleotide, what is the first corrective action made by the DNA repair system?

	The replication complex excises the incorrect nucleotide.
	The excision repair proteins remove the incorrect nucleotide.
	The mismatch repair system scans for base-pairing mismatches.
	There is no correction, because nucleotide errors by DNA polymerases are not repaired.
	DNA ligase connects the correct nucleotide.

18.

Choose the correct order of the following four events in the excision repair of DNA:
(1) Base-paired DNA is made complementary to the template.
(2) Damaged bases are recognized.
(3) DNA ligase seals the new strand to existing DNA.
(4) Part of a single strand is excised.

	1, 2, 3, 4
	2, 1, 3, 4
	3, 4, 2, 1
	2, 4, 1, 3
	4, 2, 3, 1

19.

Six complete cycles of PCR should result in a _______-fold increase in the amount of DNA.

	64
	32
	12
	128
	6

20.

When double-stranded DNA is heated to temperatures above 90Â°C, it denatures. Denaturation is a process that

	breaks covalent bonds.
	hydrolyzes DNA.
	separates double-stranded DNA into two single strands.
	causes new hydrogen bonds to form.
	irreversibly destroys the double helical structure.

Multiply questions here

Q2. What is the consequence of chromosome inversion on meiosis? (work on a zygote with one normal chromosome and one chromosome with an inversion)

Q3. Search the web or read the textbook. Find a disease that is caused by translocation. What kind of translocation causes the disease?

Q4. Briefly explain how downâs syndrome or other trisomy is generated (3 sentences should be enough for you to explain this clearly)

Q5. I mutated a culture of wild type E. coli cells and wanted to isolate mutants that are unable to synthesize uracil (a nitrogen base in RNA). How can I achieve my goal? (try to finish your answer in 2-3 sentences)

Q6. For the ura- mutant isolated in Q5, I want to convert it to URA+ by transformation. Please tell me how I can achive this transformation with 3 SIMPLE sentences

Q7. What is the difference and relationship between an Hfr cell and an F+ cell?

Q8. I want to set up a mating between an Hfr strain and an F- (ade- his- bio-) strain and detect bio+ recombinant. Please design the experiment for me. (What genotype for Hfr do you need to choose? How will you select your recombinant? etc).

Q9. I want to use phage T2 to transducer the wild type ADE2 gene from the wild type E. coli strain to an ade2- E. coli mutant. How can I do this experimentally? (please briefly describe the approach, not experimental details such as incubation for 10 min, etc).

Q10. We talked about recombination in both eukaryotes and prokaryotes. Are there any differences in these two recombination ? (what are they if there are?)

Q11. Whose (or what) experiment proves that DNA is the genetic material?

Q12. What knowledge did Watson and Crick use in constructing the DNA model?

Q13. How does the double helix model support the three major functions of genetic materials?

Q14. What is the major chemical force that holds DNA and histone together? Hydrogen bonds

Q15. If I label E. coli cells in an N15 medium overnight and chase cells in an N14 medium for 2 generations, what fraction of cells still carry the N15 label?

Q16. If a 1000 kilobase fragment of DNA has five evenly spaced replication origins and DNA polymerase moves at 1 kilobase per second, how many seconds will a cell take to produce two daughter molecules?

Q17. DNA polymerases catalyze the polymerization of nucleotides. Use the formula for enzyme catalyzed reaction (see the example) to describe DNA polymerases. Example: Enzyme Substrate A + substrate B -------------------ï product c + product D Q8. During DNA replication, what is the direction of the growing strand?

Q18. For DNA synthesis in cells, why are there a leading strand and a lagging strand?

Q19. The following is the DNA sequence of a template strand. If the primase begins to synthesize primers from the middle âgâ, what will the primer sequence be? (assuming the primer can be 8 nucleotides long) 5â GACTCTAgGACTCCT 3â

Q20. Both DNA polymerase I and DNA polymerase III synthesize DNA during DNA replication. Please state the different roles they played during DNA replication.

sangriawallaby972

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.

MBG 2040 Lecture Notes - Lecture 6: Tobacco Mosaic Virus, Ribonuclease, Deoxyribonuclease

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