DNA Modification And Repair Flashcards
Form of normal DNA modification
Methylation
What are the major methylated bases in prokaryotes?
Adenine and Cytosine
-happens after replication
N6 methyladenine
N4 methylcytosine
What is the role of methylation in bacteria?
Methylation in bacteria occurs at specific sites
Protects the bacteria’s DNA from cleavage by restriction endonucleases
What is the functional importance of methylation of adenine in E.coli?
Methylation of adenine residues in the sequence GATC is involved in mismatch error correction
Can be on either A or C or both
Cytosine in Eukaryotes
The only normal base modification in eukaryotes
5-methylcytosine
3-5% cytosine content of most DNA
Where is the methylation present in Eukaryotes?
5-methylcytosine is usually found in C residues 5’ to G
When a C in one strand is methylated, the C in the complementary strand is also methylated
What does it mean for methylation in eukaryotes to be heritable?
1) Sites of new methylation can be selected during gametogenesis by a de novo methylase
2) Not all C’s are methylated
3) fertilization and replication
4) Methylation is carried out by a maintenance methylase after replication
How does eukaryotic methylation leave room for error recognition?
Daughter strands not immediately methylated after replication –> so you can do error repair earlier on
How does methylation control gene expression?
Unmethylated promoters expressed to produce the protein
Methylated promoters are inactive and are not expressed
What is 5-azacytidine?
Inactivity of methylated genes reversed by treatment of 5-azacytidine
5-azacytidine is a cytosine analog that can be metabolized into dCTP and incorporated into DNA
How does 5-azacytidine work as a potential treatment for beta-thalassemia?
1) a normal methylated DNA goes through replication in 5-azacytidine
2) The cystosine in the daughter strands are actually 5-azacytidine
3) Replication of these new DNA leads to methylation after
4) one daughter DNA is normally methylated (due to the original methylation on that strand)
5) one daughter DNA has a loss of methylation leading to gene expression
Factors that cause DNA mutation
- Mistakes during replication
- Reactive oxygen species (ROS)
- Chemical damage
- Radiation
- ionizing agents
- Deamination of a cytosine or 5-methylcytosine
What base does a deamination of 5-methylcytosine (5mC) lead to?
It become thymine (Bc of that CH3 on C5)
Would become uracil if not methylated cytosine
How does 5-methylcytosine cause a transition mutation?
1) Deamination of methylated cytosine
2) Changed to a thymine
3) Replication of this leads to a daughter DNA which makes an originally what should be a GC base pair become an AT base pair
4) one daughter DNA is normal
Why is Deamination of 5-methylcytosine seen as dangerous if not repaired?
Thymine is not seen as an abnormal base by repair mechanisms but it’s legit changing the gene sequence that the DNA should be encoding