Module 3E – DNA Structure Objective # 25

and Replication
Summarize the evidence
 In this module, we will examine:
from the 1920s through the
 the molecular structure of the
genetic material early 1950s that convinced
 how the genetic material replicates scientists that DNA is the
 how damage to the genetic material genetic material.
is repaired
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Objective 25 Objective 25
 Scientists knew the genetic material
 During the late 19th and early 20th
centuries, scientists studying patterns must carry out 2 basic functions:
of inheritance concluded that inherited code information

traits are controlled by “hereditary replicate itself

factors” or alleles which are located on  They also knew coding could be done
chromosomes. by varying the sequence of monomers
 However, a key question remained that make up a polymer. [Similar to the
unanswered: what are these “hereditary way we code information in books by
factors” made of? varying the sequence of 26 letters.]
3 4

Objective 25 Objective 25
 Since lipids are not true polymers, and
 An important breakthrough came in
since most polysaccharides are made of
1928, when Frederick Griffith
repeating glucose units, this left 2 main
determined that something can pass
candidates: proteins and nucleic acids.
from one cell to another and alter the
 Proteins were considered the more likely
characteristics of the recipient cell.
candidate because coding information
 Griffith called this process
would be more efficient using molecules
transformation::
transformation
made of 20 different monomers rather
than just 4.
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1
 Griffths Experiment Objective 25
Mixture of Heat-Killed
Live Virulent Live Nonvirulent Heat-killed Virulent and Live  Since the agent that passed from one
Strain of Strain of Virulent Strain Nonvirulent Strains
S. pneumoniae S. pneumoniae of S. pneumoniae of S. pneumoniae cell to another in Griffith’s experiment
Polysaccharide
No
Polysaccharide
+
actually alters the characteristics of the
coat

recipient cell, many scientists believed

coat

it must be the genetic material.

 In 1944, Avery, MacLeod, and

Mice die Mice live Mice live Mice die

McCarty determined that the
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Their Lungs contain transforming agent was DNA.
live pathogenic
strain of S.
pneumoniae
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Objective 25
 In spite of this evidence, many
scientists continued to believe that
proteins, rather than DNA, function as
the hereditary material.
 The question was finally settled in
1952 by Hershey and Chase who
carried out a classic experiment with
bacteriophages:
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Objective # 26 Objective 26
 Once scientists determined that the
genetic material was composed of
Name the scientists who DNA, there was intense competition to
originally discovered the determine the structure of this
molecule
molecule.
structure off DNA.
DNA
 Scientists hoped that the structure of
DNA would provide important clues to
understanding how it works to replicate
itself and control genetic traits.
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2
 Objective 26 Chargaff’s Rules
 The structure of DNA was finally  Erwin Chargaff determined that
discovered at Cambridge University in  Amount of adenine = amount of
1953 by James Watson and Francis thymine
Crick.
 Amount of cytosine = amount of
 Their discovery was based primarily on guanine
x-ray crystallography data collected by
 Always an equal proportion of purines
Maurice Wilkins and Rosalind Franklin
(A and G) and pyrimidines (C and T)
as well as on the chemical analysis of the
base composition of DNA carried out by
Irwin Chargaff. 13 14

James Watson and Francis Crick – 1953

Rosalind Franklin
 Watson and Crick deduced the
 Performed X- X-ray diffraction studies structure of DNA using evidence from
to identify the 33--D structure Chargaff, Franklin, and others
 Discovered that DNA is helical

 Us
Usingg Maurice
au ce Wilkins’
W s DNA
DN  Did not pperform a single
g experiment
p
fibers, discovered that the themselves related to DNA
molecule has a diameter of 2 nm
and makes a complete turn of the  Proposed a double helix structure
helix every 3.4 nm

15 16

Objective # 27
Objective 27

 All nucleotides have 3 parts:

Describe the structure of a
A pentose (5-
(5-carbon) sugar
DNA nucleotide and explain
A nitrogenous
g base attached to the
h iit diff
how differs from
f the
h 1 prime carbon of the sugar
structure of an RNA A phosphate group (PO4) attached

nucleotide. to the 5 prime carbon of the sugar:

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3
 Structure of a Nitrogenous base There are 5 possible nitrogenous bases and 2
Nucleotide NH2 possible sugars:
7N 5 6
N1 Nitrogenous base
Nitrogenous Base

Phosphate group 8 7N
NH2
6
NH2 O

N C C N C C
2 5
1 H C
N
H C
N H

Purines
O 9 N 4 N3 Phosphate group
O
8

4
2 N C
N
C H N C
N
C NH2
N N
9 3
H H
Adenine Guanine
–O
P O CH2
–O P O CH2 5´
O–
5’ O NH2
C
O

C
O

C
O–
4´ 1´
H C N H3C C N H H C N H

Pyrimidines
O 3´ 2´
OH in RNA
H C
N
C O H C
N
C O H C
N
C O

OH
H H H
H in DNA
4’ 1’ Sugar Cytosine
(both DNA and RNA)
Thymine
(DNA only)
Uracil
(RNA only)

3’ 2’ OH in RNA Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

OH R
Copyright © The McGraw-Hill Companies, Inc. Permission
required for reproduction or display.
Sugar H in DNA
19 20

Objective 27 Objective # 28

 DNA nucleotides contain the sugar

deoxyribose and the nitrogenous bases Describe the structure of a
A, G, C, T.
DNA molecule and explain
 RNA nucleotides contain the sugar how it differs from the
ribose and the nitrogenous bases A, G, structure of a RNA
C, U.
molecule.
21 22

Objective 28
Double helix
 DNA:

 consistsof 2 unbranched chains of Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
5´
• each strand is a polymer 
Phosphate group of nucleotides
DNA nucleotides twisted into a double P
5´
O
• Phosphodiester 
helix..
helix backbone – repeating 
1´
4´ Phosphodiester bond
3´ 2´

P
5´
sugar and phosphate
sugar and phosphate 
O

 the 2 chains are held together by

4´ 1´ units joined by 
3´ 2´
phosphodiester bonds
hydrogen bonds between the
P
5´
O
4´ 1´
5-carbon sugar
• Nitrogenous bases 
3´ 2´
project from the sugar‐
nitogenous bases. P 5´

4´
O
1´
Nitrogenous base

phosphate backbone
3´
2´

 A always pairs with T, and G with C.

OH
3´

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4
 Complementarity of bases:
Hydrogen
bond H

H N O H N H
• G forms 3 hydrogen 
N H
bonds with C G N H N C
Sugar N N
• A forms 2 hydrogen  N H Sugar

bonds with T
bonds with T H
Hydrogen
• Gives consistent  H bond

N
diameter H N H O CH3

N A N H N T H
Sugar N N
H Sugar
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

25 26

Objective 28

 RNA is a polynucleotide composed

of a single, unbranched chain of
RNA nucleotides.

27 28

5’ end
Copyright © The McGraw-Hill Companies, Inc. Objective # 29
RNA
Permission required for reproduction or display.

P
In detail, describe the process of
Phosphate group
O
Phosphodiester
DNA replication including the
P
O
bonds names and functions of the
enzymes involved.
involved Be able to
P
O 5-carbon sugar (ribose) explain why DNA replication is
discontinuous along one strand
P O Nitrogenous base
(A,C,G or U)
and continuous along the other.
OH
3’ end 29 30

5
 Objective 29

 In 1958, Meselson and Stahl carried out

an experiment at Cal Tech supporting
the hypothesis that the replication of
DNA is semiconservative
semiconservative..
 According to this hypothesis, during
replication, the 2 strands of DNA
separate and each one serves as a
template for a new strand.
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Objective 29

 The following animation provides a

brief overview of the process of
semiconservative DNA replication.
 Note that the ability of one strand of
DNA to act as a template for the
matching strand is based on the fact
that A always pairs with T and G
always pairs with C.
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Objective 29
 The next animation provides a more
detailed description of the process of
DNA replication including the names and
functions of the major enzymes involved.
 Note that the main enzyme involved in
strand elongation, DNA polymerase III,
cannot initiate synthesis of a new
nucleotide strand. It can only add
nucleotides to the 3’ end of an existing
strand.
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6
 Objective # 30 Objective 30

 DNA replication in prokaryotes and

Explain how DNA eukaryotes is fundamentally the same.
replication in prokaryotes  On the next slide we can see how the

differs from DNA single circular DNA molecule found in

prokaryotic organisms like E. coli is
replication in eukaryotes. replicated. (Note that the circular
DNA molecule is drawn as linear in
order to simplify the animation.)
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39 40

Objective 30 Objective 30

 Inorder to speed up the replication of

 Although DNA replication in
prokaryotes and eukaryotes is very
similar, there are a few differences.
 One difference is due to the fact that
eukaryotes have so much more DNA
than prokaryotes.
41
such a large amount of DNA, the
linear DNA molecule found in each
eukaryotic chromosome may have
hundreds or even thousands of
replication origins, while the circular
DNA molecule in a prokaryotic cell
has just one.
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 Objective 30 Objective 30
 Because DNA polymerase III can only
 Another difference in the replication add new nucleotides to the 3’ end of
an existing strand, there is no way to
process is related to the fact that
complete the final segment of the
eukaryotes
y have linear rather than lagging strand located at each end of a
circular DNA. This creates a unique linear DNA molecule.
problem for eukaryotes.  As a result, linear DNA molecules gets
shorter and shorter with each round of
replication:
43 44

Original strands shown in green and newly synthesized strands in blue:

Objective 30
Eukaryotes have 2 ways to deal with
Replication first

5
round
3

3
Leading strand (no problem) Lagging strand (problem
5
this problem:
at the end)
5
3
3
5
1) The ends of eukaryotic chromosomes
Last primer
have extra DNA in the form of
Origin

Leading 3
Pi
Primer removall telomeres.. Telomeres consist of a short
telomeres
strand 5
5 nucleotide sequence that is repeated
over and over again. As a result, when
Lagging
strand 3

Removed primer
Replication second
round
cannot be replaced
eukaryotic chromosomes replicate the
5
3 5
3
telomeres shorten rather than the actual
5 3 protein--coding genes.
protein
Copyright © The McGraw-Hill Companies, Inc. Permission
3 5 46
required for reproduction or display. Shortened template

Objective 30 Objective 30
2) Some cells, such as germ cells, have  Scientists believe that normal shortening
an enzyme called telomerase.
telomerase. This of telomeres during DNA replication
enzyme has an internal RNA may help protect against cancer by
template that is used to lengthen the limiting the number of divisions that
cells can undergo.
undergo
telomeres so that chromosomes can
 Interestingly, telomerase activity has
continue to replicate without
been detected in cancer cells. By
shortening the actual protein
protein--coding lengthening the telomeres, this may
genes. allow the cells to continue to divide
47
indefinitely. 48

8
 Objective # 31

Describe the importance

and the mechanisms of
DNA repair.

49 50

Objective 31
 Many DNA polymerases can
“proofread” added bases and correct
mistakes as DNA is being replicated.
This increases the accuracy of
replication but some errors still occur
replication, occur.
 These mistakes or mutations are a
mixed blessing. They provide the
genetic variation that is essential for
evolution but, unfortunately, most are
harmful. 51 52

Objective 31
 In addition to mistakes that occur during
replication, DNA is constantly exposed to
damaging agents such as UV light, X- X-rays,
and chemicals.
 Mechanisms to repair this damage fall into
2 categories: specific and non
non--specific.
 Specific repair mechanism target a
particular type of damage. Some
examples are shown in the next animation.
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 Objective 31
 Excision repair is a non-
non-specific repair
mechanism that can be used if only
one strand of the DNA is damaged. It
involves 3 steps:
 Recognition of the damage
 Removal of the damaged strand
 Synthesis of a new strand using the
undamaged strand as a template

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