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Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Chapter 11
Key Concepts:
Biochemical Identification
of the Genetic Material
Information
Replication
Transmission
Variation
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Treatment
Result
Conclusion
1 Control:
Type S cells
are virulent.
2 Control:
Type R cells
are benign.
3 Control:
Heat-killed
type S cells
are benign.
Injected living
type S bacteria
into mouse.
Injected living
type R bacteria
into mouse.
4 Injected living
type R and
heat-killed
type S bacteria
into mouse.
Virulent type S
strain in dead
mouses blood
Living type
R cells have
been
transformed
into virulent
type S cells
by a
substance
from the
heat-killed
type S cells.
Known as transformation
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
HYPOTHESIS
A purified macromolecule from type S bacteria, which functions as the genetic material, will be able to convert type
R bacteria into type S.
KEY MATERIALS Type R and type S strains of Streptococcus pneumoniae.
Conceptual level
Experimental level
DNase
RNase
Protease
+ Type R cells
Add
antibody
Control
+ DNA
+ DNA
+ DNase
C
+ DNA
+ RNase
D
+ DNA
+ Protease
E
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Type S cells
in supernatant
Type R cells
in pellet
Centrifuge
THE DATA
B
C
DNA extract
Control
CONCLUSION DNA is responsible for transforming type R cells into type S cells.
7 SOURCE Avery, O.T., MacLeod, C.M., and McCarty, M. 1944. Studies on the Chemical Nature of the Substance Inducing
Transformation of Pneumococcal Types. Journal of Experimental Medicine 79:137156.
DNA
Protein
Phage head
(capsid)
Sheath
Tail fiber
Base plate
E. coli cell
T2 genetic
material being
injected into
E. coli
50 nm
11
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Experiment 1
Experiment 2
E. coli cells were
infected with
32P-labeled phage
and subjected to
blender treatment.
Bacterial cell
Bacterial cell
Phage DNA
32
S-labeled sheared
empty phage
P-labeled
phage DNA
35
Transfer to tube
and centrifuge.
Transfer to tube
and centrifuge.
Supernatant
has 35S-labeled
empty phage.
Supernatant has
unlabeled empty
phage.
Pellet has
E. coli cells
infected with
unlabeled
phage DNA.
Pellet has
E. coli cells
infected with
32P-labeled
phage DNA.
THE DATA
Extracellular 35S
100
Extracellular 32P
80
Blending removes 80%
of 35S from cells.
60
40
20
0
0
12
2.
3.
4.
5.
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Nucleotides
Single strand
Double helix
14
DNA
Nucleotides composed of
three components
Phosphate
Pentose
group
sugar
Base
Deoxyribose
Nitrogenous
base
Phosphate
Deoxyribose
15
DNA nucleotides
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Purines
(double ring)
Base
O
O
NH2
CH2
Phosphate
H
OH
H H
CH3
H
Thymine (T)
NH2
O
N
H
N
Adenine (A)
Deoxyribose
Pyrimidines
(single ring)
Guanine (G)
NH2
N
N
Cytosine (C)
16
RNA
Nucleotides composed of
three components
Phosphate
Pentose
group
sugar
Base
Ribose
Nitrogenous
base
Phosphate
OH
Ribose
17
RNA nucleotides
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Base
O
O
P
O
CH2
Phosphate
OH
N
H
OH
Uracil (U)
NH2
O
N
H
N
Adenine (A)
Ribose
NH2
Guanine (G)
NH2
N
N
H
Cytosine (C)
18
O
4
CH3 5
H
3
6
H
O
O
Phosphate
CH2
5
4
H
3
Thymine
2 O
O
H
OH
2
H
1
H
Deoxyribose
19
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Strands
Backbone
Bases
O
Nucleotides are
covalently bonded
CH3
Thymine (T)
H
O
5
Phosphodiester bond
phosphate group links
two sugars
Backbone formed from
phosphates and sugars
CH2
H H
1
H
H
2
H
NH2
N
O
5
Adenine (A)
N
CH2
NH2
2
H
O
5
Cytosine (C)
H
O
O
O
H
3
Phosphodiester
linkage
CH2
O
4
H
Guanine (G)
Single
nucleotide
H
N
O
CH2
5
4
Phosphate
NH2
O
H
3
OH
1
H
Sugar (deoxyribose)
ex: 5 TACG 3
20
Rosalind Franklins
X-ray diffraction results
were crucial evidence,
suggesting a helical structure
with uniform diameter
21
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22
Base-pairing
23
24
helix
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Features of DNA
5 end
Bases
3 end
Double stranded
Hydrogen bond
Antiparallel strands
Right-handed helix
Sugar-phosphate
backbone
Stabilized by H-bonding
Specific base-pairing
Sugar-phosphate
backbone
Complete turn
of the helix
3.4 nm
One nucleotide
0.34 nm
5 end
3 end
2 nm
(a) Double helix
26
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
5 end
3 end
HO
H
H H
Adenine
N
CH2
H
N
5 phosphate
P
O
CH2
Guanine
CH3
H
Thymine
CH2
H H
Guanine
Cytosine
N
CH2
O
H
3 hydroxyl
P
O
O
O
CH2
H
O
H H
H
O
H
H
P
O
O
CH2
N
H
P
O
N
O
OH
Cytosine
Hydrogen
bond
3 end
(b) Base pairing
Key Features
Two strands of DNA form a double helix.
The bases in opposite strands hydrogenbond according to the AT/GC rule.
The 2 strands are antiparallel.
There are ~10 nucleotides in each
strand per complete turn of the helix.
5 end
Chargoffs rule
A pairs
with T
G pairs with C
Keeps width consistent
GCGGATTT 3
3 CGCCTAAA 5
Antiparallel strands
One
strand 5 to 3
Other stand 3 to 5
28
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Grooves
are revealed in
the space-filling model
Major
groove
Proteins
Minor
groove
Major groove
Minor groove
Major groove
Narrower
Minor groove
29
Model
Model
Model
Semiconservative Mechanism
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Original
double helix
First round
of replication
Second round
of replication
Parental strand
Daughter strand
(a) Semiconservative mechanism. DNA replication produces
DNA molecules with 1 parental strand and 1 newly made
daughter strand.
31
Conservative Mechanism
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Original
double helix
First round
of replication
Second round
of replication
Dispersive Mechanism
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Original
double helix
First round
of replication
Second round
of replication
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Original
double helix
First round
of replication
Second round
of replication
Parental strand
Daughter strand
(a) Semiconservative mechanism. DNA replication produces
DNA molecules with 1 parental strand and 1 newly made
daughter strand.
34
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N medium
(heavy)
15
N medium
(light)
14
THE DATA
Approximate generations after transfer to 14N medium.
< 1.0
1.0
2.0
3.0
Light
Half-heavy
Heavy
Centrifuge
Meselson, M., Stahl, F., (1958) The replication of DNA in Escherichia coli,
PNAS, 44(7):67182, Fig. 4a
36
Semiconservative replication
37
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C G
C G
5
C G
A T
C G
T A
G C
G C
T A
A T
C G
A T
G C
G C
T A
A T
C G
C G
Replication
fork
A T
T
G
T
C
A
GC
G C
T A
A T
A T
T A
T A
C G
T A
Incoming
nucleotides
G C
C G
T A
Original
(template)
strand
T A
5
C G
T A
A
Newly
synthesized
daughter strand
Original
(template)
strand
3
C G
A T
C G
T A
G C
G C
T A
A T
C G
A T
G C
G C
T A
3
C G
A T
C G
T A
G C
G C
T A
A T
C G
A T
G C
G C
T A
3
5
3
(b) The products of replication
38
Molecular Mechanism
of DNA Replication
39
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Origin of
replication
Origin of
replication
Circular
bacterial
chromosome
Site where
DNA replication
ends
Origin of
replication
DNA replication
is completed.
3 DNA replication
Replication
forks
continues in both
directions.
DNA replication
is completed.
Replication
fork
Kinetochore proteins
at the centromere
Replication
fork
(a) Bidirectional replication
40
DNA helicase
Binds
DNA topoisomerase
Relives
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DNA topoisomerase
travels slightly ahead
of the replication fork
and alleviates coiling
caused by the action
of helicase.
5
DNA helicase travels
along one DNA strand
in the 5 to 3 direction
and separates the DNA
strands.
DNA polymerase
Covalently
links nucleotides
Deoxynucleoside
triphosphates
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA polymerase
catalytic site
trand
s
e
t
a
l
p
Tem
Incoming
deoxynucleoside
triphosphates
43
Deoxynucleoside triphosphates
Free nucleotides with three phosphate groups
Breaking covalent bond to release pyrophosphate (two
phosphates) provides energy to connect nucleotides
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
A
T
G
C
G
C
3 end
H
5 end
P O CH2
O
HH
O CH2
H H
HH
HH
OH
O P
O
C
H
H N
H
N H
N
H H
OH
CH2 O P
O
O
CH2
CH3
New phosphoester
bond
O
H
H N
H
N
N
H
P O CH2
O
H
OH
H
H
H
N
N
O
CH2 O P
5 end
O P O
P O
Pyrophosphate
An incoming nucleotide
(a deoxynucleoside triphosphate)
(b) Chemistry of DNA replication
H
O
3 end
N H
H
P O
O
O
CH2 O
H N
O
O
H
O
5 end
CH2 O P O
CH2 O
P O
HH
H H
H N
H
N
H H
P O CH2
N H
HO
H
P O C 2
5 end
H H
3 end
O
H
O P
CH2 O P
H
H H
H N
O
CH3
O
O
Template
strand
HO
HH
H
3 end
O
O
P
O
P O
Phosphate
2.
45
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5
RNA
primer
3
3
(b) 5 to 3 direction of
DNA synthesis
46
Leading strand
DNA synthesized
DNA primase
DNA polymerase
adds nucleotides in a 5 to 3
direction as it slides forward
Lagging strand
DNA synthesized
Okazaki
In both strands
RNA primers
47
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Replication
forks
Leading
strand
RNA primer
3
5
3
Direction of
replication fork
5
Primer
3
3
5
3
5
Second
Okazaki
fragment
First
Okazaki
fragment
5
3
5
48
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3
3
Third
Okazaki
fragment
5
3
5
49
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Origin of replication
Leading
strand
Lagging
strand
5
3
3
Replication
fork
Lagging
strand
Replication
fork
Leading
strand
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA
primase
3
5
RNA
primer
5
3
3
5
5
5
3
Second
primer
Second
primer
DNA
primase
Missing
covalent bond
3
Third
primer
Clamp
protein
3
5
Leading
strand
DNA
polymerase I
DNA
polymerase III
3
First
RNA primer
5
Lagging strand
(Okazaki
fragment)
Replicacin en E. coli
5
3
DNA ligase
3
Third
primer
52
2.
3.
DNA
DNA
DNA polymerase
DNA polymerase
DNA polymerase
When
Lesion-replicating
Telomeres
58
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3
GGG T T AGGG T T AGGG T T AGGG T T A GGG T T A
CCCAATCCCAATCCCAAT
3 overhang
5
59
61
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Telomere
5
Telomere
Eukaryotic
chromosome
3
3
Telomerase binds to a
DNA repeat sequence.
Repeat sequence
5
T
T A G G G T T A G G G T T A GG G T T A G G G T T A G GG T
A T C C CA A T
A A U C C C AAU
T A GG G T T AG G G T T AG G G T T A
AT C C CAA T
A A U C C C AA U
3
Telomerase synthesizes
a 6-nucleotide repeat
sequence.
RNA in
telomerase
Telomerase
T
T A G G G T T A G G G T T A G G G T T A GG G T
AT C C CAA T
A A U C C C AAU
T A GG G T T A GG G T T A G G GT T A G G G T T A GG G T T A
A T C C C A A T C C C A A T C C CA A U C C C A A U C C C
3
Molecular Structure of
Eukaryotic Chromosomes
would be 1 meter
Chromosome
Discrete
complex
63
DNA wrapping
DNA wrapped
Shortens
2.
30-nm fiber
Current
Shortens
64
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Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Nucleosome:
8 histone proteins +
146 or 147 nucleotide
base pairs of DNA
H2B
H2B
DNA
H2A
Linker
region
H3
H4
H4
Amino
terminal
tail of
histone
protein
30 nm
H1
11 nm
65
Interaction between
30-nm fibers and
nuclear matrix
Each chromosome
located in discrete
territory
Level of compaction is
not uniform
Heterochromatin
Euchromatin
30-nm fiber
ne
Radial loop
domain
Gene
Ge
3.
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Ge
ne
66
Cell division
not as compact
Hetrochromatin
67
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Formation of a 3-dimensional
zigzag structure via histone
H1 and other DNA-binding
proteins
11 nm
Histones
Nucleosome
Histone H1
30 nm
a: Dr. Gopal Murti/Visuals Unlimited; b: Ada L. Olins and Donald E. Olins/Biological Photo Service; c: Courtesy Dr. Jerome B. Rattner,
Cell Biology and Anatomy, University of Calgary
68
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300 nm
700 nm
(e) Heterochromatin
1,400 nm
69
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DNA double helix
2 nm
Formation of a 3-dimensional
zigzag structure via histone
H1 and other DNA-binding
proteins
11 nm
Histones
Nucleosome
Histone H1
30 nm
300 nm
700 nm
(e) Heterochromatin
1,400 nm
70