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BIO130H1 Lecture Notes - Lecture 4: Reverse Transcriptase, Germ Cell, Neuroblastoma

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maroonrat252
10 Jun 2015
School
UTSG
Department
Biology
Course
BIO130H1
Professor
Melody Neumann

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BIO130H1 Full Course Notes
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BIO130H1 Full Course Notes
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Document Summary

This course does not distinguish between dna polymerase i and iii. Steps in bacterial dna replication: origin of replication, binding of initiator proteins, unwinding by helicase, binding of single-strand binding proteins, rna primers made by primase, dna polymerase, sliding clamps holds polymerase into dna, nick sealing by dna ligase. Initiator proteins binds to origin of replication (a-t rich) Initiator proteins helps helicase that is bound to a loading protein to bind to the origin: requires atp. After initiator proteins bind for replication in e. coli: primase binds to helicase to form primosome (dna helicase + Primase: primase makes rna primer with 3" oh so dna polymerase can start producing, dna polymerase cannot add dna de novo, rest of replication machinery binds. Helicase unwind and separate strands (unzip: two types of helicase exist, the predominant one moves at 5" -> 3" along the lagging strand template (moving into the helix, helicase have 6 subunits.

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Related Questions

1) What is the difference between the leading strand and the lagging strand in DNA replication? Place the following steps of DNA replication in right chronological order.

1. Primase adds an ribonucleotide primer
2. Initiator proteins bind the origin of replication and helicase breaks hydrogen bonds between complementary bases of double stranded template DNA
3. DNA polymerase III extends by complementary base pairing deoxyribonucleotides to the template strands
4. Ligase phosphodiester bonds the nucleotides to connect okazaki fragments nad connect fragments at the origin and where the "crotches or forks" collide
5. DNA polymerase I removes RNA primer and replaces with Deoxyribonucleotides


2) Given the template DNA and primer sequences below, what bases will be added to the primer as DNA replication proceeds? (The bases should appear in the order that they will be added)

5'-GTCGTCTCCCTTTTTAAAAACCCCCCC-3'
5'-UUUUUAAAAAGGG-3'


3) Given the DNA sequence below, what could be a primer sequence used in its replication?
5'-CGTACGGCCCCAAAATTGCTACTCTGCTG-3'

Order the following steps of the DNA replication process fromfirst to last. (From 1 to 8)

DNA ligase joins Okazaki fragmments together

Elongation of leading strands occurs continuosly and elongationof lagging strands occurs via generation of Okazaki fragments. Bothare synthesized in 5' to 3' direction.

DNA polymerase I removes the RNA primers and replaces them withDNA nucleotides

DNA polymerase III starts adding DNA nucleotides to the 3' endof the existing RNA chain.

Helicase untwists the double helix at the replication fork

Single-stranded binding proteins bind the unpaired DNA strands,keeping them from re-pairing.

Topoisomerase helps releive the strain caused by DNA untwisting.It breaks, swivels and rejoins DNA strands.

Primase starts a complementary RNA chain using parental DNAstrand as a template

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.