TM-06

DNA damage, repair

and recombination

DNA REPAIR

FLASH 3. DNA Repair

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Proses perbaikan dapat dikelompokkan menjadi kategori sebagai berikut
didasarkan atas tipe kerusakan DNA:
1) Photoreactivation Repair: repairs thymine dimers caused by UV light
2) Excision Repair: repaires thymine dimers and other single strand errors
3) Proffreading and Mismatch Repair: corrects incorrect base insertion during
replication
4) Postreplication Repair: repaires incorrect base insertion after replication is
completed
5) Double Strand Break Repair: repairs breaks across both strands of a double
helix
Penjelasan lebih rinci dapat dilihat di FLASH 3. DNA Repair.

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR

Introduction
DNA molecules may acquire mutations as a
consequnce of errors in DNA replication or as
consequences of exposure to mutagens.
Living systems have evolved a variety of complex
repair systems.
Without these repair systems, mutations would
accumulate and cells (and organisms) would cease
to survive (1)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
There are a number of known repair systems.
We will cover each of these in this module.
Repair systems:
1. Photoreactivation repair
2. Excision repair
a. Base excision repair
b. Nucleotide excision rapair
3. Proofreading and mismatch repair
4. Postreplication repair
5. SOS repair
6. Double-strand break repair (2)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR

UV Radiations as a Mutagen
UV light is mutagenic, as function of the creation of
thymine dimers (two adjacent thymine molecules
become crosslinked and do not pair properly).
In order to replicate a DNA strand without error,
these dimers must be corrected. (3)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR

Photoreactivation Repair
The first important discovery concerning UV repair was by
Albert Kelner in 1949.
He showed that UV-induced damage in E. coli could be partially
reversed by exposure to visible light after irradiation.
Kelner showed that only blue light was necessary to correct
radiation damage.
The phenomenon is called photoreactivation repair. (4)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR

Blue Light and High Temperature

Kelner further showed that the blue-light effect was
temperature sensitive.
Temperature sensitivity suggests that a protein is involved
in the process (protein structure is disrupted at higher
temperature leading to loss function). (5)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR

Photoreactivation Enzyme
Photoreactivation repair is dependent on a protein
called the photo-reactivation enzyme (PRE).
The enzyme cleave the bonds between two adjacent
thymine dimers.
Although PRE can assosiate with thymine dimers in
the dark, it requires a photon of light to cleave the
dimer. (6)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Excision Repair
Excision repair is a widespread, multistep process in
which damaged DNA is removed and resynthesized
using the opposite strand as a template.
Excision repair was originally discovered because of
its ability to repair thymine dimers, but it can be
used for any damaged segment of DNA.
There are two types of excision repair:
base excision repair (BER) and
nucleotide excision repair (NER).
Both types of excision repair involve removal of
nucleotides and gap filling by DNA polymerase I. (7)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Nucleotide Excision Repair
The
1.
2.
3.

steps in nucleotide excision repair are:

The distortion or error is recognized.
The damaged area is removed by a nuclease.
DNA polymerase I fills the gap by inserting nucleotides
complementary to the other strand.
4. Ligase seals the final nick in the backbone that remains at
the 3-OH end of the base inserted.

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Base Excision Repair
Base excision repair (BER) involves an extra level of
enzyme activity prior to excision of the damaged
area.
Base alterations may result
from spontaneous hydrolysis of
a base or mutations induced by
mutagenic chemicals.
The first step in BER involves
recognition of the damaged
base by enzymes called DNA
glycosylases.
This is a group of enzymes that
are spesific to different kinds of
DNA damaged.
The enzyme cuts the bond

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Two Individuals with Xeroderma pigmentosum
Xeroderma pigmentosum (XP) is a human disease
characterized by skin lessions and multiple tumors.
Patients with XP are deficient in one of the several
genes involved in nucleotide excision repair.
Examination of their cells has helped elucidate
details of the excision repair process. (9)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Cell Fusion
Studies using a technique known as somatic cell hybridization
have been used to identify the number of different genes
involved in xeroderma pigmentos. (10)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Nucleotide Excision Repair Determined by Study
of XP
Human somatic hybrid studies have identified at least 7 genes
involved in nucleotide excision repair and have elucidated
details of the process shown here.
Remember that each of these proteins is the product of at least
one gene and a defect in any of these proteins will reduce or
eliminate nucleotide excision repair, leading to xeroderrma
pigmentosum. (11)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Correction of Error During Replication
DNA polymerase is not prefect.
The enzyme occasionally inserts an incorrect base.
DNA polymerase has the ability to reverse and
correct the error by cutting out the incorrect
nucleotide and replacing it.
This is called proofreading repair. (12)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
A Mismatch Missed by Proofreading
Even after proofreading repair, some errors persist.
To cope with these errors, a mechanism called mismatch repair
may be acivated.
A special problem exists with a mismatch.
Which strand is correct and which is the mutation? (13)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Mismatch Repair in Bacteria
At least in bacteria, the template strand is methylated and a
methyl group is added to adenine residues.
The
newly
synthesized
strand
remains
temporarily
unmethylated.
A repair enzyme binds to the unmethylated strand.
A nuclease nicks the DNA strand.
The DNA is unwound, degraded, and replaced until the
mismatch is reached and removed. (14)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Repair Occurring After Replication
Sometimes, damaged DNA escapes all repair systems. When
this happens, a final post-replication repair system may
function.
When DNA polymerase encounters a lesion such as a thymine
dimer during replication, it first stalls, then skips over the lesion
and continues, leaving a gap in the newly synthesized strand.
A protein complex, RecA, directs a recombination event from
the undamaged parent strand.
This leaves a gap in the parent strand that can be filled by the
repair process involving DNA polymerase I as seen earlier.
Post-replication repair is sometimes called homologous
recombination repair, a more general category. (15)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Undamaged DNA: SOS Repair System Repressed
Sometimes, in the bacterium E. coli, another system
exists to repair DNA damage missed by other repair
systems.
This is called the SOS repair system (after the wellknown distress call).
The SOS repair system creates conditions that allow
DNA polymerase to replicate across a damaged area.
This system is error prone and sometimes the wrong
base is inserted, a phenomenon known as SOS
mutagenesis.
A large number of gene product including LexA and
RecA are involved in this process.

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Undamaged DNA: SOS Repair System Repressed
When the LexA protein product is produced it
prevents the transcription of the recA and other
repair genes involved in the SOS repair system.
Howover, if RecA protein is present, if binds to the
single stranded DNA in the area of damage, and
activates the cleavage of the LexA repressor from the
repair genes.
When RecA binds at the site of DNA damage, the
regulatory capacity of the LexA protien is disrupted.
SOS repair genes are transcribed and their protein
products move to the repair site.
RecA protein appears to form a bridge at the lesion,
and the SOS proteins convert the lesion to an errorprone site.
This allows DNA polymerase to complete the
replication
CP 6.3:
menjelaskan across
perbaikan the
DNA gap. (16)

DNA REPAIR
Double Strand Break Repair
Some mutagenic agents create breaks across both strands of
the doule helix. Ionizing radiation (X-rays and gamma rays, for
ezample) is known to do this.
In mammals, a specialized form of repair, DNA double-strand
break repair (DSB) repairs this type of DNA damage.
The system mediates re-annaeling of the damaged DNA. (17)

CP 6.3: menjelaskan perbaikan DNA

DNA REPAIR
Homologous Recombination Repair

CP 6.3: menjelaskan perbaikan DNA

Breaks across both strands of

DNA can also be repaired by
a poorly understood process
called
homologous
recombination
repair
in
which
the
undamaged
strand.
This occurs in the late S/G2
part of the cell cycle.
This process is not well
understood.
When damage is severe, a
non-homologous portion of
the chromosome may be
used to repair the breaks.
Clearly, this process can lead
to mutations. (18)

DNA REPAIR
Module Summary | DNA Repair
1.It is clear that DNA repair is an extremely important
process and a wide variety of mechanisms exist to
ensure genetic integrity.
When these systems fail, the result is mutation
leading to abnormal cell function, cancers, and cell
death.
A number of genetic diseases are thought to be
associated with defective repair processes.
2.Repair processes can be roughly divided into the
following categories based on the type of DNA
damage that they repair:
a.Photoreactivation
Repair:
repairs
thymine
dimers caused by UV light
b.Excision Repair: repairs thymine dimers and other
single-strand errors.
c.Proofreading
Mismatch Repair: corrects
CP 6.3:
menjelaskan perbaikan and
DNA