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Lecture 12 - DNA Replicationv2

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15 views34 pages

Lecture 12 - DNA Replicationv2

Presentation with notes abput DNA replication

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itzansa
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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DNA: Genetic Material

Overview

History of DNA
DNA Structure
DNA Replication
Eukaryotic Replication
DNA Repair
Frederick Griffith

In 1928, Frederick Griffith was working on a


vaccine for the flu
At that time, they thought the cause of
the flu was a bacteria called
Streptococcus pneumoniae
During his experiment he got some
unforeseen results
There existed two strains of S. pneumoniae
S form: which was virulent and causes pneumonia
R form: non virulent, does nothing

S: Virulent R: Safe S: Virulent S: Virulent R: Safe


Griffith Experiment
Somehow the information for virulence that was
in the dead s type bacteria had passed on to the
live R type.
This transfer of information is called transformation
Bacteria pick up DNA and incorporate it into their
DNA
Avery, MacLeod, & McCarty
Experiment
 They prepared samples of Griffith’s heat killed S-Strains
and added them to different enzyme solutions
 The enzyme solutions would break down specific
biomolecules
 Dnase – breaks down DNA
 Proteinase – breaks down proteins
 Rnase – breaks down RNA
 And others
 They added the R strain like in Griffith’s experiment and
observed which would transformation
The Hershey and Chase Experiment

During this time, many scientists believed that


proteins were in charge of storing heredity
Hershey and Chase wanted to test this idea
They used a bacteriophage
Virus that only consists of DNA and a protein Coat
Two Experiments were held
DNA Structure

Once it was learned that genes were


made up of DNA, the structure of the DNA
became the next big wonder
The science community knew DNA was
made up of nitrogen bases
The purines: Adenine and Guanine
The pyrimidines: Thymine and Cytosine
Erwin Chargaff
Decided to analyze in detail the DNA of many
different species
He found that each species had different amounts of
A, T, G and C
Each species had equal amounts A = T and G = C.
For humans, we had 31% A and T and 19% G and C
His data suggested that DNA has a means to be
stable, where A can only pair with T and G can only
pair with C
Also that DNA can be variable for genetic material
Rosalind Franklin

Studied DNA using X-


ray crystallography
Her data showed
DNA to be a helix with
some portions
repeating over and
over
Types of Possible DNA Replication

There were 3 Models of replication at this time


Conservative Model: Parent Strands of DNA will still
be intact and new copies will be in the new strands
Semiconservative: Daughter strands will consist of
one parent and one new strand
Dispersive: New and old DNA is dispersed
throughout each strand of both daughter
molecules
Jump to long image description
Meselson and Stahl
Nitrogen is important for DNA replication
They grew bacterial cells in an isotope of
Nitrogen (Nitrogen 15) for many generations
After several generations, the DNA was denser
than normal DNA because of Nitrogen 15
isotope
The sample was then switched back to Nitrogen
14 and DNA samples were extracted from the
cells at different generations
Watson and Crick Model

Worked together to create a model for


DNA
Based on the previous experiments, they
knew
DNA contains 4 types of nucleotides with
bases A, T, G, and C
Amount of A = T and G = C
DNA is a double helix with repeating patterns
Watson and Crick’s Model

Created a twisting model


of DNA
Model suggested how
replication works
The model mostly holds
true today, a few
changes
They won the Nobel Prize
DNA Structure
 DNA is a double helix, like a twisted ladder
 Both strands run ANTI Parallel, meaning they run in opposite directions
 Composed of nucleotides
 5-carbon sugar called deoxyribose.
 Phosphate group (PO4).
 Attached to 5′ carbon of sugar.
 Nitrogenous base.
 Adenine, thymine, cytosine, guanine.
 Free hydroxyl group (—OH).
 Attached at the 3′ carbon of sugar.
Nucleotide subunits of DNA and RNA

Access the text alternative for slide images


© 2020 McGraw-Hill Education. 14-22
Phosphodiester bond

Bond between adjacent


nucleotides

Formed between the phosphate


group of one nucleotide and the 3′
—OH of the next nucleotide

The chain of nucleotides has a 5′-


to-3′ orientation

© 2020 McGraw-Hill Education. 14-23


Figure 11.5 DNA structure. Copyright © The McG raw-Hill Compan ies, In c. Permission re quired for repro duction or display.

C P
sugar-pho sph ate
G
C backbone
P
G
T

A P
P
A

T
P complementary
hydrog en base pairin g P
bon d
OH G
P
2' 3'
5' T
A C
S 1' 1' S 4'
4' P
3' 2' 5' P
3' end 5'end P
C
OH 5' end
a. Nucleotide pair

P G
hydrog en bonds

sugar

OH 3' end
b. Str ucture of DNA
DNA Replication

Process of copying DNA


DNA unravels, and each strand is used as a
template for the new strands
Since each new DNA molecule is made of one
parent strand, this is known to be
SEMICONSERVATIVE
DNA Replication DNA Polymerase only moves
5 prime to 3 prime
So it is a little more difficult on the
Other Strand
RNA DNA Helicase
RNA Primase
PolymeraseDNA Splits the strand of DNA
Ligase

Notice the Gap


That needs to be fixed
We need to fuse these patches of
Okazaki Fragments
RNA
Primase
DNA
Polymerase

Now it is time to create the


New To start with
strand the replication
Don’t Forget DNA Polymerase
RNA Primase comes in
Both Strands Run ANTIPARALLEL
RNA Structure
Ribonucleic Acid (RNA)
Contains sugar ribose
Base Pairs are A, C, G, and Uracil (not thymine)
Single stranded
3 types
Messenger RNA (mRNA) – used for protein synthesis
Transfer RNA (tRNA) transfers amino acids from
cytoplasm to ribosomes
Ribosomal RNA (rRNA) – creates ribosomes
Figure 11.8 Structure of RNA.
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

P
S Based in uracil
U
instead of thymine
P

S
A

G P

S C
U

A P

one S
C
nucleotide ribose
DNA Repair

DNA can become damaged by multiple different ways


UV Light, X rays, chemicals in the environment
Factors that increase the number of mutations above regular
levels of mutation are called Mutagens
DNA can be repaired usually by two different ways
Specific Repair mechanisms
Nonspecific Repair mechanisms
Photorepair: Specific repair
mechanism
When UV light hits DNA,
the strand can misshapen
in areas that have two
Thymine bases near each
other
These is called thymine
dimers
An enzyme called photolyase
uses light energy to break the
thymine dimer
Excision Repair: Nonspecific Repair
Mechanisms
A damaged region is simply excised or removed and is
then replaced
Works on 3 steps:
Recognition of damage
Removal
Resynthesis of template using other strand
Telomeres and Telomerase
There are specialized structures
on the ends of chromosomes
called telomeres
These structures protect and
maintain the integrity of the
chromosome
Consist of many short repeated
segments of DNA
Telomerase: is the enzyme that
creates and lengthens the
telomeres
Telomerase and Age

During embryonic and childhood development,


telomerase is in high activity
However, as we age, telomerase activity drops
When the telomeres shorten, the protection of
the chromosome slowly drops as well
Eventually it would lead to the non-viability of
the offspring due to the chromosomes being
damaged

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