Exam 2- DNA replication Flashcards
(T/F) Synthesis of the leading and lagging strands requires a primer only at the start of replication
false
true for leading strand but the lagging strand requires a new primer for every Okazaki fragment that is synthesized
(T/F) Base-paired primers consist of RNA that are synthesized by an enzyme
true
_______ the enzyme that synthsizes base-paired primers
DNA primase
____ used by DNA primase to synthesize short RNA primers
ribonucleoside triphosphates (ATP,CTP,GTP, UTP)
which of the ff does not apply to primers?
a. about 10 nucleotides long in eukaryotes
b. are made every 100-200 nucleotides along the lagging strand
c. has a free 3’OH group at its end used to initiate DNA synthesis
d. removed by RNAse H
none of the above, all applies :)
(T/F) In prokaryotes, Pol I removes the RNA primer and replaces it with dNTP’s
true
_____ enzyme that seals adjacent lagging strand fragments
DNA ligase
(T/F) cells use RNA primers to mark these initial (error) sequences as potentially inaccurate regions of new strands that must be replaced with correct complementary sequences
true
(T/F) enzymes capable of initiating synthesis of new DNA strands can correct their own base-pairing errors
false
would not be efficient at correcting their own base-pairing errors
______ are the proteins involved in the separation of the parental complementary strands
DNA helicases and Single-strand DNA binding proteins (SSB proteins)
____ the enzyme that functions to to pry apart double helix
DNA helicase
(T/F) DNA helicase does not require the use of energy to carry out its function
false
uses energy derived by hydrolysis of ATP to change its shape and migrate along a DNA single strand
(T/F) type of helicase that runs along the lagging strand seems to play the major role in separating the DNA strands
true
____ the structure of helicase
hexameric (6 subunits)
(T/F) do not unwind duplex but rather assist helicase by stabilizing unwound single stranded DNA
true
(T/F) aside from stabilizing the unwound DNA, helix-destabilizing proteins also function to straighten single stranded DNA of the leading strand template to prevent the formation of hairpin loops (DNA folding back on itself)
false
it does this to the LAGGING strand
____ are formed when DNA folds back on itself to self-hybridize block the movement of DNA polymerase during DNA synthesis
hairpin loops
which of the ff does not apply to single stranded binding proteins?
a. blocks hairpin formations
b. covers the bases when bound to DNA
c. prefer to bind next to previously bound molecules along a DNA strand
d. does not unwind the DNA strands
B
exposure of bases needed for base-pairing is not blocked by protein binding
_____ keeps Pol III attached to DNA during polymerization
Accessory (sliding) Clamp-Like Protein
Beta subunit of pol III in bacteria
which of the ff does not apply to sliding clamp proteins?
a. assembly requires GTP
b. its assembly is mediated by the clamp loader complex
c. Ring slides freely over single stranded DNA
d. released from DNA when it runs into DNA double strands
A
requires ATP
____ human version of E. coli clamp protein
proliferating cell nuclear antigen (PCNA)
*(T/F) in both the lagging and leading strands, Pol remains associated with clamp protein for long distance
false
this applies only to leading strand
*(T/F) on lagging strand, Pol and its associated clamp protein are released each time they reach the 5’ end of an adjacent Okazaki fragment then cycle back to the next RNA primer to synthesize another Okazaki fragment
true
____ a complex formed in prokaryotes when the helicase is directly linked to the primase on the lagging strand
primosome
(T/F) The primase is driven by the helicase along the lagging strand, producing the RNA primers needed for each successive Okazaki fragment
true
_____ the size of the major replicating protein complex
greater that 10^6 daltons in MW
(T/F) lagging strand loops back in such a way that both the leading and lagging strands can be synthesized by the same replication protein complex
true
(T/F) the pol of the of the lagging strand is associated with the large protein complex, and cannot be reused to synthesize each successive Okazaki fragment
false
it can be reused!
_____ are enzymes that regulate the supercoiling of DNA and prevent its tangling during replication (controls topology, or geometric configuration of DNA)
DNA topoisomerases
(T/F) DNA must be unwound ahead of replication fork by topoisomerases to permit movement of DNA polymerase
true
____ the speed at which the DNA helix ahead of the replication fork rotates
50 revolutions/sec
_____ the speed which the a replication fork moves
at 500 nucleotides/sec
______ the topoiomerase that produces transient single-strand breaks in the DNA along the phosphodiester backbone
type I topoisomerase
_______ the topoisomerase that generates transient double stranded breaks in DNA that can permit one DNA duplex to pass through another duplex
type II topoisomerase
(T/F) type II topoisomerase permits one strand of DNA to swivel relative to the other and thus relieve tension in the DNA during replication
false
this is type I
(T/F) DNA gyrase is situated ahead of the replication fork to unwind DNA and permit passage of DNA polymerase
true
which of the ff topoisomerases breaks the DNA strand by forming a covalent bond between a tyrosine in its active site and a DNA phosphate?
a. Type I topoisomerase
b. Type II topoisomerase
A
which of the ff topoisomerases functions to detangle chromosomes?
a. Type I topoisomerase
b. Type II topoisomerase
B
(T/F) type II topoisomerase uses energy from the hydrolysis of ATP to break permanently both strands of one DNA helix and then pass a second helix through the break
false
break transiently
(T/F) type II topoisomerase reseals the double stranded break before dissociating from the DNA
true
(T/F) DNA gyrase in bacteria are targeted as anti-tumor agents
false
as ANTIBIOTICS, does not affect eukaryotes
(T/F) humans are affected by antibiotic drugs that target DNA gyrase
false
eukaryotic cells dont produce this enzyme
(T/F) humans are affected by antibiotic drugs that target DNA gyrase
false
eukaryotic cells dont produce this enzyme
______ the species used in studies of replication in eukaryotes
yeast (S. cerevisiae)
(T/F) based on studies done on yeast, it was concluded that DNA replication in eukaryotes is very similar to that of bacteria
true
which of the ff APPLIES to prokaryotic replication?
a. SSB protein consists of one protein
b. require two or more DNA Pols
c. SSB protein is composed of three
d. Primase is a component of a multisubunit complex containing DNA polymerase alpha
A
everything else applies to eukaryotes
(T/F) On lagging strand DNA Pol alpha synthesizes an RNA primer followed by short stretch of DNA
true
(T/F) the short RNA primer synthesized by DNA pol alpha is extended into an okazaki fragment by DNA pol delta and a clamp protein
true
___ is the principal polymerase in eukaryotes
DNA pol delta
(T/F) Eukaryotes uses two DNA pols
true
alpha and delta
(T/F) in prokaryotes, , primase is associated with the DNA pol alpha
false
this is for eukaryotes!
(T/F) in bacteria, primase is associated with DNA helicase in the primosome
true
(T/F) Eukaryotic DNA is packaged in nucleosomes unlike that of bacteria
true
bacteria DNA exists as a circular DNA
(T/F) in prokaryotes, the replication fork passes through nucleosomes without dislodging them from the DNA by an unknown mechanism
false
this is in eukaryotes,
bacteria doesn’t have chromosomes
nucleosomes occur in about every ____ base pairs in eukaryotes
200
fragments are about the same size
____ the length of okazaki fragments in bacteria
1000-2000 nucleotides
(T/F) eukaryotic replication forks move 10 times slower than those of bacteria
true
Presence of nucleosomes may hinder movement of DNA Pol thats why they are slow
____ the site along a DNA molecule at which DNA replication begins
origin of replication
_____ the figure in which both parental strands are separated and are in the process of being replicated
replication bubble
which of the ff is not true about the origin of replication?
a. G-C rich
b. contains several hundred pairs long in bacteria and yeas
c. attracts initiator proteins
d. initiation mechanism is similar in bacteria and eukaryotes
A
is A-T rich coz its easier to separate
(T/F) Genome of E. coli consists of a single circular DNA molecule with several origin of replication
false
contains only one origin
(T/F) DNA replication in bacteria only only occurs in presence of sufficient nutrients to support completion of replication
true
which of the ff is the right order of proteins in the replication initiation in E. coli?
a. initiator proteins, DNA primase, primosome, DNA helicase, additional proteins
b. initiator proteins, DNA helicase, DNA primase, primosome, additional proteins
c. DNA primase, primosome, initiator proteins, DNA helicase, additional proteins
d. initiator proteins, DNA helicase, additional proteins, DNA primase, primosome, initiator proteins
B
____ the experiment performed which indicated eukaryotic chromosomes have multiple origins of DNA replication by exposing DNA in photographic emulsion
autoradiography
_____ used to radioactively label newly synthesized DNA
3H-thymidine (tritium labeled)
(T/F) DNA has bidirectional growth from a central origin of replication occurs at multiple sites along chromosomes
true
which of the ff is false about the eukaryotic origins of replication?
a. occurs in clusters
b. origins are separated by thousands of base pairs within a replication unit
c. replication forks terminate when they reach the end of the chromosomes
d. origins are in replication units
none of the above
(T/F) the RNA primers needed to produce the last Okazaki fragment cannot be removed and replaced by DNA using the normal mechanism at the 5’ end of the parental strand
false
this is the the 3’ end!
leading strand goes to the 5’ end
_____ are the ends of linear eukaryotic chromosomes
telomeres
(T/F) the lagging strand copies the telomeres to prevent shortening of DNA with each round of DNA replication
true
(T/F) At the end of a linear chromosome there is no more upstream template strand to which a primer can hybridize and initiate DNA synthesis of the 5’ end of the lagging strand
true
_____ a protein-RNA complex that recognizes and binds repeats of the 3’ end of the parental strand
telomerase
(T/F ) telomerase is a type of reverse transcriptase since it uses an RNA template to synthesize DNA
true
(T/F) the telomeres contain tandem repeats that functions to attract telomerase and are not highly conserved among all eukaryotes
false
first part is true
telomeres are highly conserved!
(T/F) Telomerase binds at the 3’ end of lagging strand and uses reverse transcriptase activity and its own RNA as a template to synthesize a DNA extension of the 5’ end of the lagging strand template with the help of RNA primer
false
binds to the 5’ of lagging strand, extends the 3’ end
(T/F) at the completion of telomere extension, the 3’ end is longer than the 5’ end with which it is paired
true
_____ is formed when telomerase-extended 3’ protruding end at each telomere becomes associated with specialized proteins
t-loop
which of the following is false about telomeres?
a. reduced telomerase activity is associated with cellular senescence
b. telomeres mechanism to regulate a cell’s lifespan
c. adult cells like cancer cells express telomerase activity
d. are expressed in highly proliferative cells
C
Most adult cells do not express telomerase
which of the ff cells do not have telomerase activity?
a. hematopoietic stem cells
b. intestinal epithelial cells
c. cells of hair follicles
d. most adult stem cells
D
_____ were used to demonstrate the involvement of telomerase in maintaining telomeres and its involvement in aging
transgenic mice
(T/F) mice telomeres are longer and needs 3 generations of breeding before its length is similar to that of humans
true
which of the ff is not seen in telomerase KO mice?
a. Defects in highly proliferative tissues
b. premature aging
c. Increased tendency to develop tumors
d. chromosomal iinstability
none of the above!
all are exhibited in KO mice
____ a disorder where humans carry one functional and one nonfunctional copy of telomerase RNA gene. symptoms include skin, lung and liver abnormalities and usually die of bone marrow failure
dyskeratosis congenita
_____ when cells withdraw from the cell cycle and cease dividing
cellular senescence
(T/F) Pol I uses its 3’ —> 5’ exonuclease activity to remove the incorrect nucleotide and then its 5’ —> 3’ polymerase activity to add the correct nucleotide
true
(T/F) sustained DNA damage, like nicks or mismatched bp can be removed by pol 1’s 5’ —> 3’ exonuclease activity similar to the mechanism of removing RNA primer
true
this works on SHORT STRETCHES/ PATCHES of DNA
(T/F) Exonuclease activity begins at a nick in the phosophodiester backbone
true
____ the mechanism by which bacteria repair base-pair mismatches that were not fixed by proofreading activities of Pol I or Pol III
mismatch repair
(T/F) in mismatch repair, any of the strands can be corrected to fix the bp mismatch
false
parental (correct strand) must be recognized by the presence of methylated bases to distinguish it from newly synthesized strand
(T/F) in E. coli, all G residues in the GATC sequence are methylated and are recognized
false
A residues are methylated
(T/F) the mismatch repair system contains MutH, MutS, and MutL that recognizes the mismatch and methylated parental strand and is the same in E.coli and in humans
false
E. coli has all three
humans lack MutH and recognizes nicks in the new strand (not methylated A)
(T/F) in E. coli, mismatch repair mechanism must operate after replication but before the new DNA strand has been methylated
true
(T/F) – People with mutations in genes that encode proteins of the mismatch repair system have a predisposition for certain types of cancers
true
*(T/F) pol I removes RNA primer and replaces nucleotides of the lagging strand at the 3’-5’ end
false
both occur 5’ to 3’ end of lagging strand
*(T/F) during DNA repair, pol I removes the incorrect nucleotide of the growing chain using its 3’ to 5’ exonuclease activity
true
_______ removes RNA primers in eukaryotes/ recognizes RNA/DNA hybrids
RNAse H
(T/F) Helicase uses energy derived by hydrolysis of ATP to change its shape and migrate along a DNA single strand; migrates in opposite polarities along the two DNA strands
true
_______ another name for single-strand DNA binding proteins
helix-destabilizing proteins
(T/F) base pairing is blocked by helix destabilizing proteins
false
_____ clusters of origin of replications
replication units