DNA REPAIR MECHANISM

SB Mirza FA11-RBI-004 sbmirza89@gmail.com

Contents
1- Introduction 2-Repair mechanisms 5- Hereditary DNA repair disorders

Introduction
DNA repair refers to a collection of processes by which a cell identifies and corrects damage to the DNA molecules that encode its genome. Sources of damage: 1-loss of a bases resulting in apurinic/apyrimidinic (AP) sites (abasic sites). 2-base modifications, such as alkylations or deamidations(hypozanthin) which converts cytosine, adenine and guanine to uracil.

3-Replication errors and base conversions can generate mismatch nucleotide pairs 5-Photodamage by uv light can generate pyrimidine dimers, 6-Chemical agents and reactive oxygen species (ROS) can modify bases .

Types of DNA Damage Summarised

G A T

C
Mismatch

ds DNA Break

C-U deamination

ss Break AP site Thymidine dimer

DNA Damage & Mutation

DAMAGE 1-Damages are physical abnormalities in the DNA 2-DNA damages can be recognized by enzymes, and, thus, they can be correctly repaired. 3-If a cell retains DNA damage, transcription of a gene can be prevented, and, thus, translation into a protein will also be blocked
MUTATION 1-A mutation is a change in the base sequence of the DNA. 2-A mutation cannot be recognized by enzymes once the base change is present in both DNA strands, and, thus, a mutation cannot be repaired. 3-Mutations can cause alterations in protein function and regulation. Mutations are replicated when the cell replicates

REPAIR MECHANISMS
In addition to DNA polymerase 35 exonuclease(the DNA Pol III has proofreading capabilities that correct replication mistakes by means of exonuclease activity working 3'->5') ..Mammalian cells utilize TWO major DNA repair pathways:

- single strand damage

Reverse Excision BER NER MMR

-db strand breaks

Non Homo Homo

A-SS damage : 1- reversal ( direct reversal ) : These mechanisms do not require a template, since the types of damage they counteract can occur in only one of the four bases.(this type repaired without removing abase or nucleotide) E.X: 1-The formation of pyrimidine dimers upon irradiation with UV light results in an abnormal covalent bond between adjacent pyrimidine bases. The photoreactivation process directly reverses this damage by the action of the enzyme photolyase.

Damage Reversal

Photoreactivation (the enzyme DNA potolyase captures energy from light )

2-Excision : In which the damaged base or bases are removed and then replaced with the correct ones . a-Base excision repair : DNA's bases may be modified by deamination or alkylation. the DNA glycosylase can recognize the damaged site and remove its base forming AP site ( Apurinic/ Apyrimidinic). Then, the AP endonuclease removes the AP site and neighboring nucleotides. The gap is filled by DNA polymerase I and DNA ligase. : -Each DNA glycosylase is generally specific for one type of lesion . _ Humans have at least four types glycosylase with different specifictices .

B- Nucleotide excision repair : NER differs from BER in several ways: -It uses different enzymes. -Even though there may be only a single "bad" base to correct, its nucleotide is removed along with many other adjacent nucleotides; that is, NER removes a large "patch" around the damage . - In NER a multisubunit enzyme hydrolyzes two phosphodiester bonds one on either side of the distorsion caused by lesion ( in human it hydrolyzes the 6th bond on 3\ side & the 22 bond on the 5\ end producing a fragment of 27-29 nucleotides ) resulting in gap filled by DNA polymerase1 & finally DNA ligase seals the nick .

UvrABC

Pol1
5 3 OR 3 5

3
5 3 5 3 5 3

3 5
3 5

5
3 5 3 3 5 3 5

3 5

3 5

C- Mismatch repair : To repair mismatched bases, the system has to know which base is the correct one. In E. coli, this is achieved by a special methylase called the "Dam methylase", which can methylate all adenines that occur within (5')GATC sequences. Immediately after DNA replication, the template strand has been methylated, but the newly synthesized strand is not methylated yet. Thus, the template strand and the new strand can be distinguished.

B- Db strand damage : There are two mechanisms by which the cell attempts to repair a complete break in a DNA molecule: 1-Direct joining: of the broken ends. This requires proteins that recognize and bind to the exposed ends and bring them together for ligating. They would prefer to see some complementary nucleotides but can proceed without them so this type of joining is also called Nonhomologous End-Joining (NHEJ). Errors in direct joining may be a cause of the various translocations that are associated with cancers.

Hereditary DNA repair disorders

Defects in the NER mechanism are responsible for several genetic disorders, including: Xeroderma pigmentosum: hypersensitivity to sunlight/UV, resulting in increased skin cancer incidence and premature aging Cockayne syndrome: hypersensitivity to UV and chemical agents Trichothiodystrophy: sensitive skin, brittle hair and nails Mental retardation often accompanies the latter two disorders, suggesting increased vulnerability of developmental neurons.

Other DNA repair disorders include: Werner's syndrome: premature aging and retarded growth Bloom's syndrome: sunlight hypersensitivity, Ataxia telangiectasia: sensitivity to ionizing radiation and some chemical agents . Huntington disease:Neurological disorder(increased number of glutamin array)