Saturday, 26 July 2014

DNA Replication

By Nur Farrah Dini

1)    Mechanism of replication
a.    Is unidirectional & semiconservative
b.    Contain one intact parental strand of each daughter DNA molecule
c.    Occurs at sites of origin of both strands which have specific base sequences
d.    Occurs only at one point of origin in each strands
2)    Steps in DNA replication include
a.    Helicases which unwind the double helix without using energy
b.    Unwinding and separating the 2 strands & form a ‘V’ where active synthesis occur
c.    Replication of unidirectional double-stranded DNA
d.    Maintaining the double helix by SSB proteins
3)    Topoisomerases enzyme
a.    Type I appear to store energy from the phosphodiester bond they sealed
b.    Irreversibly cut one strand of the double helix
c.    Create a transient ‘nick’ in one DNA strand, relieve the accumulated supercoils and resealed it
d.    In eukaryotes, DNA gyrase can be inhibited by antibiotics of quinolone family
4)    Single stranded DNA binding proteins
a.    Provides single-stranded template required by polymerases
b.    Protect DNA from nucleases that cleave single stranded DNA
c.    Serve to shift the equilibrium between double strand DNA and single strand DNA in the direction of single strand DNA forms
d.    Prevent reannealing  of the double helix
5)    The nucleotide sequences in 3’- 5’ direction
a.    Is used as the template strand
b.    Is read in that way to be copied by DNA polymerases
c.    Act as template for synthesizing the new strands of DNA in parallel direction
d.    Serves as parental strand for leading strand
6)    The synthesizing new DNA 
a.    Is initiated by DNA polymerase in both strands
b.    Requires DNA polymerase to add deoyribonucleotides to the 3’-hydroxyls of the RNA primers
c.    Involve discontinuously adding nucleotides in both strands
d.    Involve reaction of 5’triphosphate with the 3’end of the growing DNA strand forming a covalent bond
7)    Okazaki fragments are
a.    Formed in 5’- 3’direction of the leading strand
b.    Synthesized continuously in the lagging strands from 3’- 5’direction
c.    Then sealed by DNA ligase after the removal of primer & replacement of nucleotides.
d.    Formed by the DNA polymerase III after the unwinding of the double helix


Enzyme Action
Prokaryotics
a.
Leading strand synthesis
DNAP III
b.
Leading strand synthesis (RNA primer formation)
Primase
c.
Lagging strand synthesis (replacement of RNA primer with DNA)
DNAP III
d.
Joining of Okazaki fragments to lagging strands
DNA ligase
8)  
 

9)     DNA chain elongation is
a.    Adding deoxyribonucleotides randomly
b.    Done by using the 5’-hydroxyl group of the RNA primer as the acceptor of the first deoxyribonucleotide
c.    The action of DNA polymerase III adding nucleotides along the single-stranded template
d.    Result in the newly synthesized strand in the antiparallel direction to the parental strand.

10)   Proofreading of newly synthesized DNA
a.    Is done by DNA polymerase I and III
b.    Is highly important for the survival of an organism
c.    Involved activation of 5’- 3’exonuclease activity if there is any irregularity in the helix
d.    Must be done in the direction along with the newly synthesized nucleotides.
11)                        Lagging strand
a.    Synthesized continuously
b.    Being copied in the direction towards replication fork
c.    Involving small fragments of DNA being copied near replication fork
d.    Involve the joining of fragments becoming single, continuous strand
12)                        RNA primer
a.    Is formed by DNA-dependent RNA primase
b.    Consist of free hydroxyl group on the 5’end of the RNA strand
c.    Is formed for the start of synthesizing new strands
d.    Has hydroxyl group that serves as first receptor of a nucleotide
 
13)                        DNA polymerase III
a.    Includes 5’-deoxyribonucleoside triphosphates as precursors
b.    Mechanism involves releasing of pyrophosphate when each new nucleoside monophosphate is added
c.    Involves in proofreading mechanism using 5’to 3’exonuclease activity
d.    Has 3’ to 5’polymerase to replace the mistaken nucleotide with the new one
14)                        According to Francis Crick,
a.    Information flow from DNA to proteins
b.    Central dogma is well followed for most of the time
c.    Central dogma is represented by 3 main stages
15)                        Suppose a mutation occurs in a cell such that abnormal Okazaki fragments were created during DNA replication and were not linked together into a continuous strand.  The gene for which enzymes would you predict was altered by this mutation?
a.    Helicase
b.    Ligase
c.    DNA polymerase III
d.    DNA polymerase I
16)                        What happens after the RNA primer laying down a new primer on the new strands?
a.    The RNA primer is removed and is replaced by DNA.
b.    DNA polymerase I add deoxyribonucleotides
c.    Elongation from the primer producing lagging strand
d.    The 3’hydroxyls of RNA primer are being added with deoxyribonucleotides
17)                        A mutation to DNA polymerase that eliminated the 3-to-5 exonuclease activity would prevent:
a.    ligation of the okazaki fragments.       
b.    removal of the RNA primer.
c.    removal of base mismatches.        
d.     repair of deaminated bases.
18)                        The best explanation for why DNA synthesis is discontinuous would be that:
a.    DNA polymerase can only move along the DNA strand in one orientation.
b.    This allows for efficient error checking of newly synthesized DNA.
c.     DNA polymerase must stop periodically to reload more nucleotides.
d.    It happens in lagging strand
19)                        Match each of the following DNA replication mechanisms with the correct description
Mechanism                           Description
a.    DNA Polymerase proofreading            might repair a base mismatches
b.     Homologous recombination        fix a mismatch that evades DNA polymerase
c.    DNA polymerase I                                         replacement of RNA primer
d.    DNA dependent RNA primer         initiate synthesis of new strands
20)                        A mutation to DNA polymerase that eliminated the 3-to-5 exonuclease activity would prevent:
a.    ligation of the okazaki fragments.          
b.     removal of the RNA primer.
c.    removal of base mismatches.           
d.    repair of deaminated bases.
21)                        DNA replication is said to be semiconservative because:
a.     both RNA and DNA synthesis are involved in the process.
b.     part of the telomere is lost during each round of replication.
c.    a new double helix contains one old and one new strand.
d.    each new strand is complementary, not identical, to its template.
22)                        Why is an RNA primer necessary for DNA replication?
a.    The RNA primer is necessary for the activity of DNA ligase.
b.    The RNA primer creates the 5’ and 3’ ends of the strand.
c.    DNA polymerase can only add nucleotides to RNA molecules.
d.    Acts as initiator to synthesize new strands
23)                        Choose the correct statements.
a.    Single-stranded binding proteins attach after DNA helicase separates the double helix.
b.     Formation of a leading strand but not a lagging strand does not require a primer.
c.    DNA Ligase is the enzyme that links together DNA strands.
d.    RNA primers are removed and replaced with DNA before DNA ligase links together the new DNA strands.
24)                        A replication fork:
a.    is only seen in prokaryotic chromosomes.
b.    is a Y-shaped structure where both DNA strands are replicated simultaneously.
c.    is a site where one DNA strand serves as a template, and the other strand is not
replicated.
d.    is created by the action of the enzyme RNA polymerase.
25)                        Why does DNA synthesis only proceed in the 5´ to 3´ direction?
a.    Because DNA polymerases can only add nucleotides to the 3´ end of a
polynucleotide strand.
b.    Because that is the direction in which the two strands of DNA unzip.
c.    Because that is the only direction that the polymerase can be oriented.
d.    Because the chromosomes are always aligned in the 5´ to 3´ direction in the nucleus.

 Answers:

ANSWERS

A
B
C
D
1)     
F
T
T
F
2)     
F
T
F
F
3)     
F
F
T
F
4)     
T
T
T
F
5)     
T
T
F
T
6)     
F
T
F
F
7)     
F
F
T
F
8)     
T
F
F
T
9)     
F
F
T
T
10)    
T
T
F
F
11)    
F
F
T
T
12)    
T
F
T
T
13)    
T
T
F
F
14)    
T
F
T

15)    
F
T
T
F
16)    
F
F
T
T
17)    
F
F
T
F
18)    
T
F
F
T
19)    
T
F
T
F
20)    
F
F
T
F
21)    
F
F
F
T
22)    
F
F
T
T
23)    
T
F
F
T
24)    
F
T
F
F
25)    
T
F
F
F
 



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