Professional Documents
Culture Documents
218
(Molecular
Biology)
Lecture
10:
DNA
Replica9on
Dr.
John
Allingham
Bo8erell
Hall
Rm.
652
Department
of
Biomedical
and
Molecular
Sciences
allinghj@queensu.ca
February
24,
2015
We
have
covered
the
fundamental
principles
that
determine
the
informaLon-storing
and
funcLonal
capabiliLes
of
geneLc
informaLon
molecules:
We
will
now
use
these
principles
to
understand
the
molecular
basis
by
which
genomes
are
copied
and
edited,
and
geneLc
informaLon
is
regulated:
(mitosis)
(quiescent)
(four
copies
of
each
chromosome)
(each
chromosome
is
duplicated)
(two
copies
of
each
chromosome)
Some
consideraLons:
Every
Lme
a
cell
divides,
it
replicates
all
of
its
DNA;
a
process
that
takes
place
in
a
ma8er
of
hours.
Eukaryote
genomes
are
large
-
nearly
3
billion
bases
in
the
human
genome!
Considering
that
the
human
body
houses
about
15
million,
million
cells,
each
one
harboring
a
(near)
perfect
copy
of
the
genome
(two
copies
in
fact),
it
is
important
that
mistakes
are
minimized.
5
Some
consideraLons:
A
large
number
of
factors
are
associated
with
the
process
of
DNA
replicaLon
to
ensure
rapid
and
faithful
reproducLon
of
the
geneLc
material.
[32P] dTTP
incorporation
11
12
RadioacLvity
13
Results
PuricaLon
of
Polymerase
by
Kornberg
et
al.
involved
a
mulLstep
process
in
which
protein
concentraLon
and
specic
enzyme
acLvity
(DNA
polymerizaLon)
were
monitored.
(Lehman, I. R., Bessman, M. J., Simms, E. S., and Kornberg, A. (1958) J. Biol. Chem. 233, 163170)
14
Results
(Bessman, M. J., Lehman, I. R., Simms, E. S., and Kornberg, A. (1958) J. Biol. Chem. 233, 171177)
Conclusions:
DNA
Polymerase
ReacLon
Components
Requirements:
All
DNA
polymerases
require
a
template
strand.
All
DNA
polymerases
require
a
primer
strand.
Deoxyribonucleoside
5-triphosphates
(dNTPs)
Polymerase
enzyme
+
associated
cofactors
16
More consideraLons:
18
19
20
DNA polymerases
Viral
Bacterial
Human
Viral
21
23
29
KEY CONVENTION
30