Finals - DNA Repair Mechanisms Flashcards
Different DNA repair mechanisms
- direct repair
- excision repair
- mismatch repair
- double-stranded break repair
- SOS response
- involves chemical reversal of the damage without breaking the phosphodiester backbone of the DNA
- not dependent on a template since the damage does not alter the sequence within which it occurs.
Direct repair
where direct repair happens
- nicks
- alkylation damage
- cyclobutyl dimers
sinlge-strand breaks in DNA where a phosphodiester bond is missing
nicks
repairs nicks
DNA ligase
what does DNA ligase do
glues phosphodiester bonds
- repaired through enzymatic transfer of alkyl group from nucleotide to their own polypeptide chains
- removed by ADA
alkylation damage
ADA
Adenosine deaminase
Ex of alkylation damage repair
- ADA enzyme of E. coli
- Human MGMT
MGMT
methyl guanine – DNA methyl transferase
what does ADA do
- removes methyl and puts it on its cystein residue
- alkylated base is free from alkyl
what does human MGMT do
interacts with alkylating agents during chemotherapy
- repaired by DNA photolyase
- need presence of light
cyclobutyl dimers
what repairs cyclobutyl dimers
DNA photolyase
- perhaps the best known DNA lesion affecting a single DNA strand
- it is an intrastrand cross-link in which two adjaent pyrimidines are connected by a cyclobutane ring
pyrimidine dimer (PD)
where pyrimidine dimer most frequently form
two thymines (thymine dimer)
Steps in direct repair of cyclobutyl dimers
- Photolyase needs to be activated by UV. Chromophore abosorbs UV light and energy is transfered to FADH (noncovalent bonding).
- FADH’s electron is transferred to pyrimidine dimer causing it to split
- Restores and hydrogen bonds are formed (renaturation)
UV wavelength in activation of photolyase
320-370nm
wavelength chromophore absorbs
300-500nm
involves excision of a single damaged base, followed by resynthesis
base excision repair
Enzymes in base excision repair
- DNA glycosylase
- AP endonuclease
- DNA polymerase β
- DNA ligase
- involved in the removal of damaged base
- creates AP site
DNA glycosylase
what does DNA glycosylase create
apurinic/apyrimidinic site (AP site)
incise posphodiester backbone adjacent to AP site
AP endonuclease
- fill the gap created during base excision repair
- incorporate nucleotide
DNA polymerase β
seals the backbone during base excision repair
DNA ligase
Summary of Base Excision Repair
- DNA glycosylase removes damaged base and creates apurinic/apyrimidinic site (AP site)
- AP endonuclease incise phophodiester backbone adjacent to AP site
- DNA pol β fill the gap by incorporating nucleotide
- DNA ligase seals nick
- repairs damage affecting longer strands, 2-30 bases
- used by the ell for bulky DNA damage
Nucleotide excision repair
length of damage during nucleotide excision repair
2-30 bases
nucleotide excision repair is used by the cell for what?
bulky DNA damage
mediated by gene products of uvrA, uvrB, or uvrC
NER in bacteria
- involves XPA, XPB, XPC, XP6 proteins
- CSA and CSB proteins
- ERCC7, RPA, and RAD 23 proteins
NER in eukaryotes
inherited condition characterized by an extreme sensitivity to ultraviolet radiation (UVR), which is present in sunlight and may also be found in some types of artificial lighting
Xeroderma pigmentosum
what is absent in people with Xeroderma pigmentosum
Nucleotide excision repair (NER)
Summary of Nucleotide Excision Repair
- uvrB with uvrA scans damaged DNA
- uvrA helps uvrB recognize damaged part
- uvrA released, uvrC attaches, helicase II unwinds
- uvrC with uvrB excise the damaged part and some nucleotides near are also cut
- uvrB bridges the gap
- DNA pol I incorporate complemetary nucleotides
- DNA ligase seal nicks
Nucleotide excision repair (NER):
scans damaged DNA
uvrB with uvrA