GENA RELICATION

Review of NUCLEIC ACIDS

DNA
Genetic material
In the nucleus
Double stranded
A,T
C,G

The sides of the ladder are sugar

and phosphates; the rungs of the
ladder are the nitrogen bases
A,T,C,G

RNA
Carries information from the
nucleus to the site where
proteins are made
Single stranded
A,U
C,G

How DNA is the Master Control

DNA Replication
The structure of DNA provided an
insight to Watson and Crick for how
DNA replicates
each strand in a duplex of DNA is
complementary to each other,
each can form a template when
separated.
The order of bases on one strand
can be used to add in
complementary bases and
therefore duplicate the pairs of
bases exactly.
Model for DNA Replication
Watson and Cricks model:
semiconservative replication

DNA is found in the nucleus

Humans have 23 pairs of
chromosomes
Chromosomes are made of DNA

DNA molecul

Three models of DNA replication

Experiment which supports the

Semiconservative Model
Matthew Meselson and Franklin Stahl
experiments supported the
semiconservative model
labeled the nucleotides of the old
strands with a heavy isotope of
nitrogen (15N) while any new
nucleotides would be indicated
by a lighter isotope (14N).
Replicated strands could be
separated by density in a
centrifuge.
Each model:

Meselson-Stahl experiment

Conclusion: DNA replication

follows semiconservative model
Conservative and Dispersive
models were disproven in their
experiment.

DNA Replication
More than a dozen enzymes and
other proteins participate
E. coli can replicate 4.5 x 106
base pairs bp) in less than an
hour
human cells can replicate 6 x 109
bp in only a few hours
DNA replication is very accurate
less than 1 error per billion
nucleotides!!
DNA Replication Start Sites
Where does DNA replication start?
special sites termed origins of
replication
single site in bacterial
chromosome
multiple sites in eukaryotic
chromosome
Enzymes (helicases) separate the two
strands
forms a replication bubble
other proteins (single strand
binding proteins - ssb) bind to
keep strands separated
Origins of Replication

Enzymes of DNA Replication

DNA polymerases
synthesize DNA by adding a
nucleotide that is complementary to
the base in the template strand
Rate of synthesis
Bacteria - 500 nucleotides / sec
Human cells - 50 nucleotides /
sec
Incorporation of a nucleotide

DNA Synthesis
leading strand is synthesized
continuously
lagging strand is synthesized
discontinuousl in short segments
called Okazaki fragments DNA ligase
joins the fragments

Bacterial DNA Replication Proteins

Helicase
Unwinds parental double helix at
replication forks
DNA double helix are tightly
coupled. High temperature is
needed to break them (95oC

ssb protein
binds to and stabilizes ssDNA
Topoisomerase
Corrects overwinding ahead of
replication forks
breaks, swivels, and rejoins DNA
strands
Primase
synthesizes single primer for
leading strand
synthesizes RNA primer for each
lagging strand
DNA pol III
continuous synthesis of leading
strand
discontinuous synthesis of
lagging strand
DNA pol I
removes primer (RNA) from DNA
strand and replaces it with DNA
DNA Ligase
joins 3 end of fragment with 5 end
of adjacent fragment
DNA replication Fork

Summary of DNA Replication

Initiating DNA Synthesis

After separation of the DNA strands
DNA Polymerase cannot initiate DNA
syn.
Needs a 3 OH to add nucleotide to.
synthesizing a new chain requires a
primer, a short segment of RNA
Primase (an RNA Polymerase) adds
about 10 nucleotides complementary
to template
Priming DNA Synthesis
Note: RNA primer is removed from DNA
by another DNA Polymerase

Bacterial DNA Replication Proteins

Mutation
Mutation: A change in the base
sequence of the DNA
Mutations are changes in the
genotype which may or may not
affect the phenotype
Causes of mutations
Spontaneous mutations
Occur in the absence of mutation
causing agents
Due to occasional mistakes in
DNA replication

Induced mutations
Caused by mutagens, agents such as
chemicals and radiation which induce
mutations
Types of Repair of DNA
Bases may be damaged by chemical
and/or physical agents
UV light, reactive chemicals,
radiation, etc.
Some mismatched bases may be
missed by proofreading activity of
DNA pol
Must be corrected to ensure high
fidelity of DNA sequence
Types of mutation
Base substitutions
The most common type of
mutation
A single base pair is replaced by
another
Frame shift mutations
One or more base pairs are
inserted or deleted in the DNA
Results in a change in the reading
of codons
mutagen
Chemical mutagens
Example: Nitrous acid alters adenine
such that it pairs with cytosine
instead of thymine

Radiation
Ionizing radiation e.g.,

Xrays and gamma rays

Causes the formation of ions

that can react with
nucleotides (causing base
changes) and the
deoxyribose-phosphate
backbone (causes
chromosomes to break).
UV radiation
Induces formation of covalent
bonds between adjacent
thymines to form thymine dimers
which can not be replicated

Consequences of base substitutions

Silent mutation:
base change results in no change
of the amino acid sequence of
the translated protein
Silent mutations have no effect
on phenotype
A result of the fact that multiple
codons can code for the same
amino acid
E.g., AGU and AGC both code for
Serine
Missense mutation:
base change results in the
change of an amino acid in the
translated protein
The amino acid substitution
induced by the missense
mutation may have no effect on
the function of the protein OR
It may abolish the activity of the
protein or alter its function
having an effect on phenotype

Example: sickle cell disease in

humans is due to a missense
mutation in the gene for globin.
As a result the shape of red blood
cells is altered affecting their
movement through capillaries.

and the production of inactive

protein

Nucleotide Excision Repair

In nucleotide excision repair, a
nuclease cuts out a segment of a
damaged strand.
The gap is filled in by
DNA polymerase and
ligase.

Penyakit mutasi
Xeroderma Pigmentosum
Individuals with this genetic disease
have defective repair enzymes
cant remove thymine dimers
caused by UV light
very sensitive to sunlight and
often get skin cancers

Nonsense mutation:
base change generates a stop
codon in place of that coding for
an amino acid
Results in production of a
truncated protein. Usually results
in a non-functional protein

Frameshift mutation:
addition or deletion of one or
more bases
Results in misreading of the
codons (changed reading frame)
Almost always results in long
stretches of altered amino acids

Royal Hemophilia
X linked
Queen Victoria (Great Britain)
was the carrier
Gene passed on to many other
royal families in Europe

Russian, Prussian and

Spanish affected

But not the British royal

family

Genetic warfare?
Translation: Making Proteins
A group of three nucleotides in
messenger RNA codes for a certain
amino acid to be placed in a protein.
Each group of three nucleotides is
called a CODON.
Stop kodon UAG, UGA, UAA
Start kodon AUG
DNA Repair
Spontaneous DNA damage
Pathways to remove DNA damage
Damage detection
The repair of Double-strand break
DNA repair enzymes

Now you know---How DNA is the

Master Control

TAC CGA TCG

ATG GCT AGC
transcripsi
TAC CGA TCG
AUG GCU AGC new RNA
translasi
TAC CGA TCG
AUG GCU AGC met, ala, ser
So how do you remember the
difference?
Replication just makes a repeat copy
of DNA
Transcription rips a new strand of
RNA
Translation starts with a new slatethe slate of amino acids to make a
protein