Section 3

DNA Replication
Process

Replication is a continuous process , to describe

it clearly, we separate it into three stage :
Initiation: recognize the origine point,
separate dsDNA, primer synthesis,
Elongation: add dNTPs to the existing
strand, form phosphodiester bonds,
correct the mismatch bases, extending
the DNA strand,
Termination: stop the replication.

a. Initiation
Dna B
Dna A

Dna C
primase

3'

5'
3'
DNA topomerase

5'
SSB

Primosome complex

b. Elongation
dNTPs which is complementary to template are
continuously connected to the primer or the nascent
DNA chain by DNA-pol III.
The nature of the chain elongation is the series
formation of the phosphodiester bonds.

Replication Fidelity
Replication based on the principle of
base pairing is crucial to the high
accuracy of the genetic information
transfer.
Enzymes use two mechanisms to
ensure the replication fidelity.
Proofreading and real-time correction
Base selection

c. Termination
The replication of E. coli is
bidirectional from one origin, and the
two replication forks must meet at
one point called ter at 32.
All the primers will be removed, and
all the fragments will be connected
by DNA-pol I and ligase.

3'

5'
3'

5'

RNAase
3'
5'

OH
dNTP

3'
5'

5'

DNA polymerase
P

ATP
3'

5'
3'

5'
3'

DNA ligase
5'
3'

3.2 Replication of Eukaryotes

DNA replication is closely related with cell
cycle.
Multiple origins on one chromosome, and
replications are activated simultaneously.

Telomere
The terminal structure of eukaryotic
DNA of chromosomes is called
telomere.
DNA sequence is rich in TTAGGG
repeat sequence.
The telomere structure is crucial to
keep the termini of chromosomes.

Telomerase
The eukaryotic cells use telomerase to
maintain the integrity of DNA telomere.

Reverse transcription

Reverse transcription is a process in

which ssRNA is used as the template
to synthesize dsDNA.

Reverse transcriptase
Reverse transcriptase is the enzyme
for the reverse transcription. It has
activity of three kinds of enzymes:
RNA-dependent DNA polymerase
RNase
DNA-dependent DNA polymerase

Section 5
DNA Damage and Repair

5.1 Mutation
Mutation is a change of nucleic acids in
genomic DNA of an organism. The
mutation could occur in the replication
process as well as in other steps of life
process.

5.2 Causes of Mutation

R
P

N
O

O
O

UV
N

CH3

O
CH3

CH3
O

CH3

N
O

O
N

(TT)

Mutation caused by chemicals

Carcinogens can cause mutation.
Carcinogens include:
Food additives and food
preservatives; spoiled food
Pollutants: automobile emission;
chemical wastes
Chemicals: pesticides; alkyl
derivatives; -NH2OH containing
materials

5.3 Types of Mutation

a. Point mutation (mismatch)
Point mutation is referred to as the
single nucleotide alternation.
5'
3'

C T T C A G G A

3'

G A A G T C C G G C G

5'

 Transition: the base alternation from

purine to purine, or from pyrimidine
to pyrimidine.
Transversion: the base alternation
between purine and pyrimidine, and
vise versa.

Transition mutation

Hb mutation causing Sickle-cell of anemia

Single base mutation leads to one AA
change, causing disease.
HbA

HbS

chains

CTC

CAC

mRNA

GAG

GUG

AA residue 6 in chain

Glu

Val

Acidic AA Glu change to Non-polar and hydrophobic AA

When PO2 decreased, HbS molecular

interaction, a helical polymer solubility is low,
causes RBCs to sickle cell. The shape of the
RBCs was very irregular, large number of thin,
elongated, sickle-shaped forms.

b. Deletion and insertion

Deletion: one or more nucleotides are
deleted from the DNA sequence.
Insertion: one or more nucleotides
are inserted into the DNA sequence.
Deletion and insertion can cause the
reading frame shifted.

Frame-shift mutation
Normal
5 GCA GUA CAU GUC A
Ala Val His Val
Deletion C
5 GAG UAC AUG UCA
Glu Tyr Met Ser

c. Rearrangement
It is the exchang or transfer of genetic
information between homologous
chromosome ,resulting in the formation
of new characteristics not found in
either parental DNA.

Gene Rearrangement lead to mediterranean anemia

Hb gene family on
Chr 11

5.4 DNA Repairing

DNA repairing is a kind response made
by cells after DNA damage occurs,
which may resume their natural
structures and normal biological
functions.
DNA repairing is a supplementary to the
proofreading-correction mechanism in
DNA replication.

Light repairing

R
P

N
O

O
O

UV
N

CH3

O
CH3

CH3
O

CH3

N
O

O
N

(TT)

Excision repairing
Recognise and cleave
injured fragment by
endonuclease

Excision repairing
One of the most important and
effective repairing approach.
UvrA and UvrB: recognize and bind
the damaged region of DNA.
UvrC: excise the damaged segment.
DNA-pol : synthesize the DNA
segment to fill the gap.
DNA ligase: seal the nick.

Recombination repairing
It is used for repairing when a large segment
of DNA is damaged.

SOS repairing
It is responsible for the situation that
DNA is severely damaged and the
replication is hard to continue.
If workable, the cell could be
survived, but may leave many errors.