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BIOL 2030 Lecture Notes - Lecture 12: Frederick Griffith, Martha Chase, Reverse Transcriptase

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rubycheetah358
3 May 2018
School
Dalhousie University
Department
Biology
Course
BIOL 2030
Professor
Multiple

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Molecular Basis of Heredity
February 1st, 2016
DNA as a source of Genetic Material
• Replicates accurately
• Stores large amounts of information
• Allows for phenotypic variation (traits)
Historical Background:
• Jogann Meischer – isolated a weakly acidic substance…
• Frederick Griffith – demonstrated that cells could be transformed
o Virulent – kills mouse
o Nonvirulent – does not kill mouse
o Heat killed virulent type – does not kill mouse
o Mixture of nonvirulent and heat-killed virulent – kills mouse virulent bacteria
recovered **transformation demonstrated
o The nonvirulent R cells had somehow acquired the ability to cause disease
o Nonvirulent R cells had been transformed
o A transforming factor is present in the heat-killed virulent S cells
o Substance responsible..
▪ Is take up by the R strain
• Avery MacLeod and McCarthy – first demonstration that DNA was the genetic material
o Heat killed virulent S strain and extracted cell content
o Treat cell extract with virulent S strain with enzymes
o Result: transformation of R to S strain
o Conclusion: only DNase destroyed the transforming substance, th e activity is not
destroyed by RNase or protease
• Alfred Hersey and Martha Chase: DNA not protein is transmitted…..
o Life cycle of a phage: virus that infects bacteria
▪ Injects DNA within bacterial chromosome
▪ Phage = 50% protein; 50% DNA
o Experiment:
▪ Bacteria infected with phage that has 35S labeled coat protein
▪ Bacteria infected with phage that has 32P labeled DNA
▪ Separate empty phage protein coat from bacterial cells
▪ Results after phage reproduction:
• 35S – labeled protein not detected in progeny
• 32P – labeled DNA detected in progeny
▪ Therefore, DNA not protein has been transmitted to the progeny
• Heinz Frakenl-Conrat and Bea Singer – demonstrated that RNA can serve as a genetic
material
o Separated proteins from RNA (type A and type B)
o Type A RNA + types B protein (& via versa)
o This creates hybrid TMV = fully active
o Virus particles produced then match the type of RNA
o Retrovirus – RNA DNA via reverse transcriptase
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Related Questions

magine that the year is 1928 and the scientific world is in an uproar trying to determine what component of the cell isresponsible for genetic inheritance. Is it DNA, protein, perhaps even RNA? You are busy in your lab, trying to help solve this crucial mystery that still plagues (then) modern day biology. Your work focuses on a microorganism, a bacterium named Streptococcus pneumoniae, and its effects on mice. There are two strains of S. pneumoniae (which is responsible for pneumonia), the “Rough” strain (R) and “Smooth” strain (S). The strains are very similar except the S strain has a smooth polysaccharide coat that enables the microorganism to avoid the organism’s immune system and cause illness (and death in mice). The R strain has a rough coat which is detected and dealt with by the immune system (and as a result causes no death in mice). You decide to perform a series of experiments using the R and S strain of S. pneumoniae involving injections of these strains in mice. Your results are documented in the figure below: living S strain of S. pneumoniae mouse dies of infection living R strain of S. pneumoniae mouse lives mouse lives S strain heat-killed living R strain mouse dies of infection living, pathogenic S strain recovered S strain heat-killed From these results, you figure out that something from the heat killed S strain is transforming the R strain into the virulent strain. What is this “Transforming principle”? Is it related to the cellular component that was passed from one generation to the next? As you know, this was the initial experiment performed by Dr. Frederick Griffith that eventually led to the discovery of the true cellular component that is the genetic inheritance molecule as well as the “transforming principle” (Hint: It’s DNA!).

1. Several experiments were required to demonstrate how traits are inherited. Which scientist or team of scientists first demonstrated that cells contain some component that can be transferred to a new population of cells and permanently cause changes in the new cells?

a. Griffith b. Watson and Crick c. Avery, MacLeod, and McCarty d. Hershey and Chase

2. From your results, which one of the cellular components can be determined NOT to be the component responsible for genetic inheritance? Why?

a. Proteins because these macromolecules become denatured with heat. b. DNA because these macromolecules form a single helix in solution. c. RNA because it's double stranded 100% of the time d. Sugars because glycolysis happens.

3. Which one of the following experimental trials below are considered to be controls for this experiment? a. S Strain grown in 10% ethanol medium. b. Living S strain; Living R strain; Heat Killed S Strain c. Living R Strain with heat killed S strain; Living S Strain d. Heat Killed S strain; Living R strain; R strain grown in 10% ethanol medium

4. Which scientist or team of scientists obtained definitive results demonstrating that DNA is the genetic molecule? a. Griffith b. Watson c. Crick d. Hershey and Chase

5. Hershey and Chase used radiolabeled macromolecules to identify the material that contains heritable information. What radioactive material was used to track DNA during this experiment?

a. 3H b. 14C c. 35S d. 32P

6. DNA molecules, like proteins, consist of a single, long polymeric chain that is assembled from small monomeric subunits.

a. True b. False

7. The polarity of a DNA strand results from the polarity of the nucleotide subunits.

a. True b. False

8. There are five different nucleotides that become incorporates into a DNA strand.

a. True b. False

9. Hydrogen bonds between each nucleotide hold individual DNA strands together.

a. True b. False

1. 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.
2. 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
3. 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
4. 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.
5. 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
6. The uniform diameter of the DNA structure provides evidence for
antiparallel DNA strands.
a right-handed helix.
a double-stranded helix.
3′ end phosphorylation.
purine–pyrimidine pairing.
7. If a sequence of one strand of DNA is 5′-TGACTATC-3′, what is the complementary strand?
5′-ACTGATAG-3′
3′-TGACTATC-5′
3′-ACTGATAG-5′
3′-ACTGATAG-3′
5′-TGACTATC-3′
8. What structural aspect of the DNA facilitates dissociation of the two DNA strands for replication?
3′ end phosphorylation
Purine–pyrimidine pairing
Antiparallel DNA strands
Covalent bonds between sugars and phosphate groups
Hydrogen bonding between paired bases
9. If the Meselson–Stahl 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 3′ hydroxyl group (–OH). 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 90°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.