Genetics Assignment

S ubmitted to - Dr. A tik a C ha nd ra

v S ubmitted by - S wa rna l i

Rol l N o.- 19 / 14 6 3

Progra m N a me- B.S c . Bota ny ( Hons)

S emester- 3 rd semester

Ti tl e of Pa per - Geneti c s ( C C - 7 )
DNA REPAIR
MECHANISM
(Mis-match Repair)
 DNA
❖ In the cytoplasm of the cell is present DNA.

❖ DNA is a molecule that contains all the genetic

information.

❖ DNA is made up of nucleotides

each of which contains a Phosphate-
Sugar backbone and an organic base.

❖ The two strands of DNA are held together by H-

bonding with complementary base pairs, I.e.,
Adenine pairs with Thymine and Guanine with
Cytosine.
 ➢ DNA is a highly stable molecule that is replicated
with amazing accuracy but changes in
DNA structure and errors of replication do occur.

➢ DNA damage occurs during R eplication.

DNA Damage ➢ The damage can be caused by environmental
agent such as radiations, chemicals etc.

➢ The rate of mutation remains remarkably low,

thanks to the efficiency with which DNA is
repaired.
Types of DNA Damage
❖ All four of the bases in DNA (A, T, C, G) can be covalently modified at
various positions.

❖ Mismatches of the normal bases because of failure of proofreading

during DNA replication.

❖ Breaks in the backbone.

❖ Crosslinks or covalent linkages can be formed between bases.

 Damage Repair
❑ DNA damage, if not repaired, may affect replication and
transcription, leading to mutation or cell death.

❑ DNA repair is a collection of processes by which a cell

identifies and corrects damage to the DNA molecules
that encode its genome
 Common DNA Repair mechanism
Repair System Types of Damage Repaired

Mismatch Replication errors, including mis paired bases and strand

slippage.

Direct Pyrimidine dimers; other specific types of alterations.

Base Excision Abnormal bases, modified bases and pyrimidine dimers.

Nucleotide Excision DNA damage that distorts the double helix, including abnormal
bases, modified bases, and pyrimidine dimers.

Homologous recombination Double- strand breaks

Non- homologous end joining Double- strand breaks

Mis-Match Repair (MMR)

D NA mismatch re pair (MMR) is a highly conse rve d b iological pathway.

plays a k e y role in maintaining ge nomic stab ility.

The spe cificity of MMR is primarily for b ase -b ase mismatche s and
inse rtion/d e le tion mis pairs ge ne rate d d uring D NA re plication and
re comb ination.

I nactive mismatch re pair will cause spontane ous mutation.

Mismatch re pair pre ve nt mutage ne sis and tumorige ne sis.

E.coli MutS and MutL and the ir e uk aryotic homologs, MutSα and MutL α,
re spe ctive ly, are k e y playe rs in MMR associate d ge nome mainte nance .
o In E.coli, proteins involved are
1. MutS, MutL, MutH, DNA helicase ll (MutU/UvrD)
2. Exonucleases
3. Single stranded DNA binding proteins (SSB)
4. DNA polymerase lll holoenzyme
5. DNA ligase

o MutS recognizes base- base mismatches.

o MutL interacts physically with MutS, enhances mismatch recognition, and recruits
and activates MutH.

o MutH, an enzyme that causes an incision or nick one strand near the site of the
mismatch.
 Cont..
➢ Before the replication begin s, the
Helicase en zy me n eeds to un w in d the
DNA molecule.

➢ In E.coli, DNA poly merase cataly zes

the in corporation of n ew
complemen tary bases to the n ew
DNA stran d. This is on ly the alpha(α)
an d beta(β) un its of the DNA
poly merase l l l en zy me.
 Cont..

➢ It is possible for the

DNA poly merase l l l to corporate
a n on - complemen tary base to the
n ew stran d. This is k n ow n
as a Mis-match.

➢ A mis - match could result in

a faulty protein bein g coded
for so it n eeds to be repaired.

Insertion of non-
complementary base- Mis
match
 Cont..
➢ The first step in the MMR pathway of E.coli
is recognition of the mis- match by the MutS
homodimer.

➢ MutS recognizes the mis- match and bind to

it.

➢ The MutL Homodimer then binds to

the MutS.

➢ MutL interacts physically with MutS,

enhances mismatch recognition, and recruits
GATC site
and activates MutH.

➢ MutH protein recognizes a CATC site about

a 1000bp away.
 Cont..

➢ MutH, an endonuclease creates an incision in the

backbone between the guanine and adenine
residue on the newly synthesized strand.

➢ MutH differentiates between newly synthesized

strand and the template strand by the lack of
methylation markers on the newly synthesized
strand, which is why the pathways is also known
as methyl directed MMR.

➢ The mismatch-repair complex brings

the mismatched bases close to the
methylated GATC sequence, and the new strand
is identified.
 Cont.. Newly formed backbone
incision

➢ Exonucleases remove nucleotides on

the new strand betw een the GATC
sequence and the mismatch.

➢ DNA Polymerase lll begins to

resynthesize the new strand
inserting the correct complementary
base w here the mis match used to be,
 Cont..
➢ Finally, DNA ligase ligates the backbone
nick.
➢ The Mis match pathway is now
completed and all the newly synthesized
DNA is complementary to the template
strand.
➢ Mismatch repair in eukaryotic cells is similar to that in E.coli, but how the
old and new strands are recognized in eukaryotic cells is not known.

➢ Mismatch excision has been demonstrated in vitro with nuclear extracts

prepared from human cells. Thus, mismatch repair is probably a universal or
nearly universal mechanism for safeguarding the integrity of
genetic information stored in double -stranded DNA.

➢ Humans who possess mutations in mismatch-repair genes often exhibit

elevated somatic mutations and are frequently susceptible to colon cancer.
 References

• Book- pierce- genetics- conceptual- approach- 4 th

• Book- Principle of Genetics

• Li, GM. Mechanisms and functions of DNA mismatch repair. Cell

Res 18, 85–98 ( 2008) . http s://d oi.org /10.1038/cr.2007.115

• https://www.creative - diagnostics.com /mismatch - repair- pathway.htm