Cap 11 Biologia

Chapter 11
Lecture and
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Chapter 11
Nucleic Acid Structure, DNA Replication,

and Chromosome Structure
Key Concepts:
Biochemical Identification of the Genetic Material
Nucleic Acid Structure
An Overview of DNA Replication
Molecular Mechanism of DNA Replication
Molecular Structure of Eukaryotic Chromosomes

2
Biochemical Identification
of the Genetic Material
What is the genetic material?
Four criteria necessary for genetic material:

1.
2.
3.
4.
Information
Replication
Transmission
Variation
Late 1800s biochemical basis of heredity postulated
Researchers became convinced that chromosomes carry

the genetic information
1920s to 1940s scientists expected the protein portion of

chromosomes would turn out to be the genetic material
3
Griffiths bacterial transformation
Late 1920s Frederick Griffith was working with

Streptococcus pneumoniae bacteria
Two strains of S. pneumoniae:
Strains that secrete capsules look smooth (S)

and infections are fatal in mice
Strains that do not secrete capsules look rough (R) and

infections are not fatal in mice
The capsule shields the bacteria from the immune

system, so they survive in the blood
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.
Injected heatkilled type S

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.
Smooth strains (S) with capsule are fatal; rough

strains (R) without capsule are not
If mice are injected with heat-killed type S, they

survive (because bacteria are dead)
However, mixing live R with heat-killed S kills

the mouse
Blood is found to contain living type S bacteria
Known as transformation
How is this possible?
Genetic material had been transferred

from the heat-killed type S bacteria to the living
type R bacteria
This gave them the capsule-secreting trait and

was passed on to their offspring
What was the biochemical basis of this

transforming principle? At the time there was no
way to know
7
Avery, MacLeod, and McCarty Used

Purification Methods to Reveal That
DNA is the Genetic Material
1940s interest in finding biochemical basis of bacterial

transformation
Only purified DNA from type S could transform type R
But, purified DNA might still contain traces of

contamination that may be the transforming principle
Added DNase, RNase and proteases
RNase and protease had no effect
When DNase was added, no transformation took place
Surprising conclusion: DNA is the genetic material
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
Purify DNA from a type S strain.

This involves breaking open cells
and separating the DNA away from
other components by
centrifugation.
DNase
RNase
Protease
+ Type R cells
Mix the DNA extract with type R

bacteria. Allow time for the DNA
to be taken up by the type R cells,
converting a few of them to type S.
Also, carry out the same steps but
add the enzymes DNase, RNase, or
protease to the DNA extract, which
digest DNA, RNA, and proteins,
respectively. As a control, dont add
any DNA extract to some type R cells.
Add
antibody
Control
Add an antibody, a protein made by

the immune system of mammals,
that specifically recognizes type R
cells that havent been
transformed. The binding of the
antibody causes the type R cells to
aggregate.
A
+ DNA
+ DNA
+ DNase
C
+ DNA
+ RNase
D
+ DNA
+ Protease
E
Remove type R cells by

centrifugation. Plate the remaining
bacteria (if any) that are in the
supernatant onto petri plates.
Incubate overnight.
Type S cells
in supernatant
Type R cells
in pellet
Centrifuge
THE DATA
B
C
DNA extract
Control
DNA extract + RNase

DNA extract + DNase
DNA extract + protease
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.
Hershey and Chase
1952 studied a T2 virus that infects Escherichia coli
Bacterial virus is known as bacteriophage or phage
Phage coat made entirely of protein
DNA found inside capsid

DNA
DNA
Protein
Phage head
(capsid)
Sheath
Tail fiber
Base plate
E. coli cell
T2 genetic
material being
injected into
E. coli
(a) Schematic drawing of

T2 bacteriophage
50 nm
(b) An electron micrograph of T2 bacteriophage

infecting E. coli
Eye of Science/Photo Researchers, Inc.
11
Experiment 1
E. coli cells were

infected with
35
S-labeled phage
and subjected to
blender treatment.
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
Using a Geiger counter,

determine the amount of
radioactivity in the supernatant.
Geiger (radioisotope)
counter
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
Total isotope in supernatant (%)
Sheared empty phage
Extracellular 35S
100
Extracellular 32P
80
Blending removes 80%
of 35S from cells.
60
40
Most of the 32P (65%)

remains with intact cells.
20
0
0
Agitation time in blender (min)
12
Nucleic Acid Structure

Levels of DNA Structure:
1.
Nucleotides the building blocks of DNA and RNA
2.
Strand a linear polymer strand of DNA or RNA
3.
Double helix the two strands of DNA
4.
Chromosomes DNA associated with an array of

different proteins into a complex structure
5.
Genome the complete complement of genetic

material in an organism
13
Nucleotides
Single strand
Double helix
DNA associates with

proteins to form a
chromosome.
14
DNA
Formed from nucleotides (A, G, C, T)
Nucleotides composed of
three components
Phosphate
Pentose
group
sugar
Base
Deoxyribose
DNA = Deoxyribonucleic Acid
Nitrogenous
base
Phosphate
Purines Adenine (A), Guanine (G)
Pyrimidines Cytosine (C), Thymine (T)
Deoxyribose
15
DNA nucleotides
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
(a) DNA nucleotide
Pyrimidines
(single ring)
Guanine (G)
NH2
N
N
Cytosine (C)
16
RNA
Formed from nucleotides (A, G, C, U)
Nucleotides composed of
three components
Phosphate
Pentose
group
sugar
Base
Ribose
RNA = Ribonucleic Acid
Nitrogenous
base
Phosphate
Purines Adenine (A), Guanine (G)
Pyrimidines Cytosine (C), Uracil (U)
OH
Ribose
17
RNA nucleotides
Base
O
O
P
O
CH2
Phosphate
OH
N
H
OH
Uracil (U)
NH2
O
N
H
N
Adenine (A)
Ribose
(b) RNA nucleotide
NH2
Guanine (G)
NH2
N
N
H
Cytosine (C)
18
Nucleotide numbering system
Sugar carbons are 1 to 5

Base attached to 1 carbon on sugar
Phosphate attached to 5 carbon on sugar
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
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)
Bases project away from

backbone
Written 5 to 3
Single
nucleotide
H
N
O
CH2
5
4
Phosphate
NH2
O
H
3
OH
1
H
Sugar (deoxyribose)
ex: 5 TACG 3
20
Solving the structure of DNA
1953, James Watson and Francis Crick, proposed the

structure of the DNA double helix
Watson and Crick used Linus Paulings method

of working out protein structures using simple
ball-and-stick models
Rosalind Franklins
X-ray diffraction results
were crucial evidence,
suggesting a helical structure
with uniform diameter
21
X-rays diffracted by DNA

onto photographic plate
Pattern represents the
atomic array in wet fibers
Wet DNA fibers

X-ray beam
22
Base-pairing
Erwin Chargoff analyzed base composition

of DNA from many different species
Results consistently showed

amount of adenine (A) = amount of thymine (T)
amount of cytosine (C) = amount of guanine (G)
23
24
Watson and Crick
Put together these pieces of information
Found ball-and-stick model consistent with

data
Double-stranded
Base-pairing:
helix
A with T and G with C
James Watson, Francis Crick, and Maurice

Wilkins awarded Nobel Prize in 1962
Rosalind Franklin had died and the Nobel Prize

is not awarded posthumously
25
Features of DNA
5 end
Bases
3 end
Double stranded
Hydrogen bond
Antiparallel strands
Right-handed helix
Sugar-phosphate
backbone
Bases on the inside
Stabilized by H-bonding
Specific base-pairing
~10 nts per helical turn
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
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
Complementary DNA strands

5
GCGGATTT 3
3 CGCCTAAA 5
Antiparallel strands
One
strand 5 to 3
Other stand 3 to 5
28
Grooves
are revealed in
the space-filling model
Major
groove
Proteins
bind to affect gene

expression
Minor
groove
Major groove
Minor groove
Major groove
Narrower
Minor groove
29
An Overview of DNA Replication
Late 1950s three different models were

proposed for DNA replication
Semiconservative
Conservative
Dispersive
Model
Model
Model
Newly-made strands are daughter strands
Original strands are parental strands

30
Semiconservative Mechanism
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
Original
double helix
First round
of replication
Second round
of replication
(b) Conservative mechanism. DNA replication produces 1 double

helix with both parental strands and the other with 2 new
daughter strands.
32
Dispersive Mechanism
Original
double helix
First round
of replication
Second round
of replication
(c) Dispersive mechanism. DNA replication produces DNA

strands in which segments of new DNA are interspersed
with the parental DNA.
33
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.
(b) Conservative mechanism. DNA replication produces 1 double

helix with both parental strands and the other with 2 new
daughter strands.
(c) Dispersive mechanism. DNA replication produces DNA

strands in which segments of new DNA are interspersed
with the parental DNA.
34
Meselson and Stahl experiment
In 1958, Matthew Meselson and Franklin Stahl

devised an experiment to differentiate among the three
proposed DNA replication mechanisms
Nitrogen comes in a common light form (14N) and

a rare heavy form (15N)
Grew E. coli in medium with 15N to label, then switched

to medium with 14N, collecting samples after each
generation
Original parental strands would be 15N while newly

made strands would be 14N
Conclusion: Semiconservative DNA replication

35
Grow bacteria in 15N

media.
Transfer to 14N media and

continue growth for <1,
1.0, 2.0, or 3 generations.
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
Isolate DNA after each generation. Transfer

DNA to CsCl gradient, and centrifuge.
DNA
CsCl gradient
Centrifuge
Observe DNA under UV light.
Meselson, M., Stahl, F., (1958) The replication of DNA in Escherichia coli,
PNAS, 44(7):67182, Fig. 4a
36
Semiconservative replication
The two parental strands separate and serve

as template strands
New nucleotides must obey the AT/GC rule
End result: two new double helices with same

base sequence as original
37
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
(a) The mechanism of DNA replication
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
Origin of replication provides an opening

called a replication bubble that forms two
replication forks
DNA replication proceeds outward from forks
Bacteria have single origin of replication
Eukaryotes have multiple origins of replication
39
Origin of
replication
Origin of
replication
Circular
bacterial
chromosome
DNA strands unwind.
DNA strands unwind,

and DNA replication
begins.
Site where
DNA replication
ends
Origin of
replication
DNA strands unwind,

and DNA replication
begins at multiple
origins of replication.
DNA replication
is completed.
DNA replication begins outward

from two replication forks.
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
(b) Single origin of replication in bacteria
(c) Multiple origins of replication in eukaryotes
40
DNA helicase
Binds
to DNA and travels 5 to 3 using

ATP to separate strand and move fork
forward
DNA topoisomerase
Relives
additional coiling ahead of

replication fork
Single-strand binding proteins

Keep
parental strands open to act as

templates
41
DNA topoisomerase
travels slightly ahead
of the replication fork
and alleviates coiling
caused by the action
of helicase.
Single-strand binding proteins

coat the DNA strands to prevent
them from re-forming a double helix.
3
5
DNA helicase travels
along one DNA strand
in the 5 to 3 direction
and separates the DNA
strands.
Direction of replication fork

5
42
DNA polymerase
Covalently
links nucleotides
Deoxynucleoside
triphosphates
DNA polymerase
catalytic site
trand
s
e
t
a
l
p
Tem
(a) Action of DNA polymerase
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
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
Features of DNA polymerase

1.
2.
DNA polymerase cannot begin synthesis

on a bare template strand
Requires a primer to get started
DNA primase makes the primer from RNA
The RNA primer is removed and replaced with

DNA later
DNA polymerase only works 5 to 3
45
DNA polymerase is able to

covalently link nucleotides
together from a primer, which
is made by DNA primase.
DNA polymerase can link

nucleotides only in the
5 to 3 direction.
5
RNA
primer
(a) Need for a primer
3
3
(b) 5 to 3 direction of
DNA synthesis
46
Leading strand
DNA synthesized
DNA primase
in as one long molecule
makes a single RNA primer
DNA polymerase
adds nucleotides in a 5 to 3
direction as it slides forward
Lagging strand
DNA synthesized
Okazaki
5 to 3 but as Okazaki fragments
fragments consist of RNA primers plus DNA
In both strands
RNA primers
are removed by DNA polymerase and

replaced with DNA
DNA ligase joins adjacent DNA fragments
47
DNA strands separate at an

origin of replication, creating
2 replication forks.
Replication
forks
Primers are needed to initiate

DNA synthesis. The synthesis
of the leading strand begins in
the direction of the replication
fork. In the lagging strand, the
first Okazaki fragment is made
in the opposite direction.
Leading
strand
RNA primer
3
5
3
Direction of
replication fork
5
Primer
3
The leading strand elongates,

and a second Okazaki fragment
is made.
3
5
First Okazaki fragment

of the lagging strand
3
5
Second
Okazaki
fragment
First
Okazaki
fragment
5
3
5
48
The leading strand continues

to elongate. A third Okazaki
fragment is made, and the first
and second are connected
together.
3
5
3
3
Third
Okazaki
fragment
First and second Okazaki

fragments have been
connected to each other.
5
3
5
49
Origin of replication
Leading
strand
Lagging
strand
5
3
3
Replication
fork
Lagging
strand
Replication
fork
Leading
strand
(b) Replication from an origin

50
DNA primase makes RNA primers to begin

the replication process.
DNA
primase
3
5
DNA polymerase III continues to elongate

the leading strand. In the lagging strand,
DNA polymerase III synthesizes DNA
from the second primer. DNA polymerase
I removes the first primer and replaces it
with DNA.
RNA
primer
5
3
3
5
5
5
3
Second
primer
DNA polymerase III makes DNA from the

RNA primers. DNA primase hops back to
the opening of the fork and makes a second
RNA primer for the lagging strand.
Direction of replication fork

DNA
polymerase III
5
3
Second
primer
DNA
primase
Missing
covalent bond
3
Third
primer
Clamp
protein
3
5
Leading
strand
In the lagging strand, DNA ligase forms a

covalent bond between the first and second
Okazaki fragments. A third Okazaki
fragment is made. The leading strand
continues to elongate.
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
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52
DNA replication is very accurate
Three mechanisms for accuracy

1.
Hydrogen bonding between A and T,

and between G and C is more stable than
mismatched combinations
2.
Active site of DNA polymerase is unlikely to form

bonds if pairs mismatched
3.
DNA polymerase can proofread to remove

mismatched pairs
DNA polymerase backs up and digests linkages
Other DNA repair enzymes as well

53
DNA Polymerases Are a Family of

Enzymes With Specialized Functions
Important issues for DNA polymerase are speed,

fidelity, and completeness
Nearly all living species have more than one type of

DNA polymerase
Genomes of most species have several DNA polymerase

genes due to gene duplication
Independent genetic changes produce enzymes with

specialized functions
E. coli has 5 DNA polymerases

DNA
polymerase III multiple subunits,

responsible for majority of replication
DNA
polymerase I a single subunit, rapidly

removes RNA primers and fills in DNA
DNA
polymerases II, IV and V DNA repair and

can replicate damaged DNA
DNA polymerases I and III stall at DNA damage
DNA polymerases II, IV and V dont stall but go slower

and make sure replication is complete
Humans have 12 or more DNA polymerases

Designated
with Greek letters
DNA polymerase
its own built in primase subunit
DNA polymerase
and extend DNA at a faster rate
DNA polymerase
replicates mitochondrial DNA
When
DNA polymerases , or encounter abnormalities they

may be unable to replicate
Lesion-replicating
polymerases may be able to synthesize

complementary strands to the damaged area
Telomeres
Series of short nucleotide sequences repeated

at the ends of chromosomes in eukaryotes
Specialized form of DNA replication only in

eukaryotes in the telomeres
Telomere at 3 does not have a complementary

strand and is called a 3 overhang
58
Telomere repeat sequences
3
GGG T T AGGG T T AGGG T T AGGG T T A GGG T T A
CCCAATCCCAATCCCAAT
3 overhang
5
59
DNA polymerase cannot copy the tip of the

strand with a 3 end
No
place for upstream primer to be made
If this replication problem were not solved,

linear chromosomes would become
progressively shorter
Telomerase enzyme attaches many copies

of DNA repeat sequence to the ends of
chromosomes
60
Shortening of telomeres is correlated with

cellular senescence
Telomerase function is reduced as an

organism ages
99% of all types of human cancers have

high levels of telomerase
61
Telomere
5
Telomere
Eukaryotic
chromosome
3
3
Telomerase moves 6 nucleotides

to the right and begins to make
another repeat.
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
Primase makes an RNA primer near the end of

the telomere, and DNA polymerase synthesizes
a complementary strand in the 5 to 3 direction.
The RNA primer is eventually removed.
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
RNA primer that is 5

eventually removed
Molecular Structure of
Eukaryotic Chromosomes
Typical eukaryotic chromosome may be

hundreds of millions of base pairs long
Length
But
would be 1 meter
must fit in cell 10-100m
Chromosome
Discrete
unit of genetic material
Chromosomes composed of chromatin

DNA-protein
complex
63
Three levels of DNA compaction

1.
DNA wrapping
DNA wrapped
Shortens
2.
around histones to form nucleosome
length of DNA molecule 7-fold
30-nm fiber
Current
model suggests asymmetric, 3D zigzag of

nucleosomes
Shortens
length another 7-fold
64
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
(a) Micrograph of a 30-nm fiber
30 nm
H1
11 nm
(b) Three-dimensional zigzag model

a: Photo courtesy of Dr. Barbara HamkaloZ
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
Protein fiber inside the nucleus
Radial loop
domain
Gene
Radial loop domains
Ge
3.
Ge
ne
Protein that attaches the base

of a DNA loop to a protein fiber
66
Cell division
When cells prepare to divide, chromosomes

become even more compacted
Euchromatin
not as compact
Hetrochromatin
much more compact
Metaphase chromosomes highly compacted
67
DNA double helix

2 nm
(a) DNA double helix
Wrapping of DNA around

histone proteins
Formation of a 3-dimensional
zigzag structure via histone
H1 and other DNA-binding
proteins
11 nm
Histones
Nucleosome
(b) Nucleosomes (beads on a string)
(c) 30-nm fiber
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
300 nm
Anchoring of radial loop

domains to the nuclear matrix
Further compaction of radial

loops to form heterochromatin
Metaphase chromosome with

2 copies of the DNA
(d) Radial loop domains
700 nm
(e) Heterochromatin
1,400 nm
(f) Metaphase chromosome

d: Courtesy of Paulson, J.R. & Laemmli, U.K. James R. Paulson, U.K. Laemmli, The structure of histonedepleted
metaphase chromosomes, Cell, 12:81728, Copyright Elsevier 1977; e-f: Peter Engelhardt/
Department of Virology, Haartman Institute
69
DNA double helix
2 nm
(a) DNA double helix
Wrapping of DNA around

histone proteins
Formation of a 3-dimensional
zigzag structure via histone
H1 and other DNA-binding
proteins
Anchoring of radial loop

domains to the nuclear matrix
Further compaction of radial

loops to form heterochromatin
Metaphase chromosome with

2 copies of the DNA
11 nm
Histones
Nucleosome
(b) Nucleosomes (beads on a string)
Histone H1
(c) 30-nm fiber
30 nm
300 nm
(d) Radial loop domains
700 nm
(e) Heterochromatin
1,400 nm
(f) Metaphase chromosome

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; d: Courtesy of Paulson, J.R. & Laemmli, U.K. James R. Paulson, U.K. Laemmli, The structure of histonedepleted
metaphase chromosomes, Cell, 12:81728, Copyright Elsevier 1977; e-f: Peter Engelhardt/Department of Virology, Haartman Institute
70

Cap 11 Biologia

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Cap 11 Biologia

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Chapter 11

See separate PowerPoint slides for all figures and

Nucleic Acid Structure, DNA Replication,

Biochemical Identification of the Genetic Material

Nucleic Acid Structure

An Overview of DNA Replication

Molecular Mechanism of DNA Replication

Molecular Structure of Eukaryotic Chromosomes

What is the genetic material?

Four criteria necessary for genetic material:

Late 1800s biochemical basis of heredity postulated

Researchers became convinced that chromosomes carry

1920s to 1940s scientists expected the protein portion of

Griffiths bacterial transformation

Late 1920s Frederick Griffith was working with

Two strains of S. pneumoniae:

Strains that secrete capsules look smooth (S)

Strains that do not secrete capsules look rough (R) and

The capsule shields the bacteria from the immune

Injected heatkilled type S

Smooth strains (S) with capsule are fatal; rough

If mice are injected with heat-killed type S, they

However, mixing live R with heat-killed S kills

Blood is found to contain living type S bacteria

How is this possible?

Genetic material had been transferred

This gave them the capsule-secreting trait and

What was the biochemical basis of this

Avery, MacLeod, and McCarty Used

1940s interest in finding biochemical basis of bacterial

Only purified DNA from type S could transform type R

But, purified DNA might still contain traces of

Added DNase, RNase and proteases

RNase and protease had no effect

When DNase was added, no transformation took place

Surprising conclusion: DNA is the genetic material

Purify DNA from a type S strain.

Mix the DNA extract with type R

Add an antibody, a protein made by

Remove type R cells by

DNA extract + RNase

DNA extract + protease

Hershey and Chase

1952 studied a T2 virus that infects Escherichia coli

Bacterial virus is known as bacteriophage or phage

Phage coat made entirely of protein

DNA found inside capsid

(a) Schematic drawing of

(b) An electron micrograph of T2 bacteriophage

E. coli cells were

Using a Geiger counter,

Total isotope in supernatant (%)

Sheared empty phage

Most of the 32P (65%)

Agitation time in blender (min)

Nucleic Acid Structure

Nucleotides the building blocks of DNA and RNA

Strand a linear polymer strand of DNA or RNA

Double helix the two strands of DNA

Chromosomes DNA associated with an array of

Genome the complete complement of genetic

DNA associates with

Formed from nucleotides (A, G, C, T)