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DNA Replication

what constitutes a nucleotide? deoxyribose/phosphate?


nitrogenous bases? how many bonds exist between nitrogenous
bases? which are purines and which are pyrimidines (single ring or
double ring)? Identify antiparellel structure, be able to label 5' and
3' on each strand
DNA by Watson & Crick & Iranklin
\hen discoered, the structure suggested how
DNA was able to replicate:
u 1he l-bonds between complementary bases
break
u 1his allows the DNA to unzip
DNA by Watson & Crick & Iranklin
u Lach DNA strand then acts as a template to
build the complementary strand
u 1his results in two identical DNA molecules, one or
each daughter cell
1he big question one time was the method o
DNA replication, was it:
1, Conseratie: the original double stranded DNA was consered
as one molecule or one daughter, and a completely new one or
the other daughter
2, Semi-conseratie: the original molecule is split in hal, and the
other side is illed-in,
3, Dispersie: Lach new molecule is comprised o bits and pieces o
both new DNA and the original strand
Models of DNA Replication
conservative
semiconservative dispersive
J
2
P
Meselson and Stahl
u grew .cbericbia coti ,. coti., in
15
N
media, which made the DNA
heay.
u 1hen grew it on normal media
,
14
N,.
u 2 possible outcomes:
1, New molecules hae medium
,hybrid, DNA or
2, One molecule has heay DNA the
other light,
u Only the irst was obsered
1hat process is called
Semiconservative Replication
Overview.
DNA replication has many steps, but the general
order o steps is:
a, Initiation and unzipping.
b, Priming
c, Llongation
d, Proo-reading.
Initiation and Unzipping
u Replication begins when proteins bind at a
speciic site on the DNA known as the origin o
replication ,ori,.
u Lukaryotic replication is similar to prokaryotic
replication but more complex
u 1he closed circular DNA o prokaryotes usually
only has one origin o replication ,ori,
u Linear eukaryotic DNA has multiple ori`s
DNA Strand Separation
u 1he strands cannot simply be pulled apart
because they are held together by hydrogen
bonds and twisted around each other in the
double helix.
u Speciic enzymes
work together to
expose the DNA
template strands
DNA Strand Separation
u \hat is inoled
u DNA helicase: unwinds the double helix by
breaking the l-bonds at the replication ork.
Replication fork:
region where
enzymes replicating
DNA bind to an
untwisted, single
stranded DNA
strand.
DNA Strand Separation
u Single-stranded
Binding Proteins
(SSBs) bind the
exposed DNA
template strands to
block new l-bonds
that would re-join
the strands
DNA Strand Separation
u DNA gyrase: reliees tension rom the
unwinding o the DNA strands. It cuts nicks in
both strands o DNA, allowing them to swiel
around one another and then resealing the cut
strands.
Replication
u Replication begins in 2 directions rom the ori`s
as a region o the DNA is unwound.
u DNA replication proceeds in the 5` to 3`
direction
u replication moes toward the direction o the
replication ork ,leading strand, on one strand
and away rom the ork ,lagging strand, on the
other strand.
Priming
u Once the replication bubble is
started, then primase lays-
down short-sequence RNA
primers as complements to the
DNA strand.
u RNA primers are synthesized
by primase and are temporary
u 1he leading strand ,one primer, is synthesized
continuously
u 1he lagging strand ,multiple primers, is
synthesized discontinuously in short ragments
Llongation
u DNA polymerase III builds the complimentary
strand o DNA by adding complimentary
nucleotides in the 5` to 3` direction, using RNA
primers as starting points
u 1he RNA primers are needed because DNA
Polymerase III needs a 3`Ol to add to.
Elongation
1he lagging strands produces many small
segments called Okazaki fragments
Elongation
u DNA polymerase I remoes the RNA primers
rom the leading strand and ragments rom the
lagging strand and replaces them with the
appropriate deoxyribonucleotides.
Text page 221, figure 7b
Elongation
u On the lagging strand, DNA ligase joins the Okazaki
ragments into one continuous strand o DNA
Elongation
As the 2 new strands o DNA are synthesized, 2 double
stranded DNA molecules are produced that
automatically twist into a helix.
Lditing & proofreading DNA
u 1000 bases,second ~ lots o typos!
u DNA polymerase I
u prooreads & corrects typos
u repairs mismatched bases
u remoes abnormal bases
u repairs damage
throughout lie
u reduces error rate rom
1 in 10,000 to
1 in 100 million bases
Iast & accurate!
u It takes . coti 1 hour to copy
5 million base pairs in its single chromosome
u diide to orm 2 identical daughter cells
u luman cell copies 6 billion bases & diide into
daughter cells in only ew hours
u remarkably accurate
u only ~1 error per 100 million bases
u ~30 errors per cell cycle
protein RNA
1he Central Dogma
DNA
transcription translation
replication
u llow o genetic inormation in a cell