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3e DNA
3e DNA
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
1 2
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
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.
5 6
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
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:
9 10
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.
11 12
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
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
17 18
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
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
4´
O
1´
Nitrogenous base
phosphate backbone
3´
2´
23 24
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
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
Objective 29
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.
35 36
6
Objective # 30 Objective 30
39 40
Objective 30 Objective 30
7
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
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
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.
53 54
9
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
55 56
57
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