dna replication Flashcards
dna pol III
carries out genomic replication and DNA repair functions
note: high polymerization rate, high processivity, and proofreading
dna pol I
has roles in replication, recombination, and DNA repair
note: 3’->5’ proofreading and 5’->3’ exonuclease activities (repair)
pol I and pol III have _______________________
high fidelity= they don’t make a lot of mistakes
mutations
permanent errors in replication that will be inherited by subsequent daughter cells
what are the 3 mechanisms to avoid mistakes during replication?
presynthetic error control, proofreading, and mismatch repair
presynthetic error control
demands correct base pairing
presynthetic= before phosphodiester bond formation
geometry of watson and crick base pairs allows them to fit into the catalytic site of DNA polymerase
mispaired base pairs are excluded
errors can occur due to spontaneous tautomerism
proofreading
postsynthetic error control
recognizes and corrects mistakes
high-fidelity DNA polymerases have 2 active sites: catalytic site for DNA synthesis & 3’->5’ exonuclease site for proofreading
prokaryote DNA pol I, II, and III are all high-fidelity enzymes
tautomers
tautomeric base pairs result from chemical rearrangements of the bonds within the base
- occur spontaneously
- can participate in forming alternative base pairs
- isomerization of the tautomeric base to a normal base results in a mismatched base pair
bidirectional synthesis
replication forks proceed in both directions, away from the origin of replication
DNA synthesis begins at origin of replication
yields two DNA molecules with identical nucleotide sequences
replication fork
where parent DNA is being unwound (by helicase) and the separated strands are replicated by DNA polymerase
leading strand
primed only once
synthesis occurs continuously from 5’-> 3’
lagging strand
synthesis occurs discontinuously from 5’->3’
okazaki fragments
each fragment must be primed
fragments must later be joined
what is true about replication in prokaryotes?
occurs only once per cell cycle
initiation is the regulated step in control of DNA synthesis
oriC (origin of replication)
has binding sites for DnaA protein
DNA unwinding element (DUE) is an AT-rich segment where strand separation occurs (energy requirement is much lower and hydrogen bonds are easier to break)
what causes localized strand separation at the DUE (dna unwinding element)? (DNA replication initiation in prokaryotes)
binding of DnaA proteins to the R and I sites stresses the DNA helix
DnaC loads a DnaB helicase at both ends of the bubble on what will be the lagging strand. What does this lead to?
loading of DnaB helicase commits the cell to replication and division
DnaB helicase leads the replication fork and separates DNA strands 5’-> 3’
what happens after DnaC is released?
DNA pol III’s are assembled onto both DnaB helicases
DnaA is then released from the DNA
DNA pol III is ready to replicate the strand
holoenzyme
core polymerase + accesory subunits
core polymerase (pol III)
catalyzes DNA synthesis
there are 2 - one for leading strand and one for lagging strand
keeps lagging and leading moving together
clamp loader serves as…
-a scaffold for DNA polymerase III complex
-to assemble the B clamp onto DNA using ATP
-coordinates the replication fork by interacting with DnaB helicase through tau subunits
B clamp (aka sliding clamp)
tethers the core polymerase to DNA
- decreases the polymerase dissociation from DNA -> increases processivity
primase
DNA template-dependent and primer-independent RNA polymerase
- synthesizes a RNA primer at the beginning of the leading strand and each okazaki fragment
loading the DNA pol III on B clamp on DNA
B clamp encircles DNA- provides means for high processivity and rapid DNA polymerization
-ATP binding to the clamp loader causes it to open and bind to the clamp
-once open, it can associate with the DNA strand
- the clamp loader will add the clamp at site of the primer
-ATP hydrolysis causes the clamp to close around DNA
what generates the leading strand?
DNA synthesized as a continuous polynucleotide chain by one core polymerase
how is the lagging strand synthesized?
as a series of okazaki fragments
_________ functions at the replication fork just as core polymerase near completion of an okazaki fragment
- _________ begins synthesis of a new RNA primer
primase