DNA Replication & Repair Flashcards
(38 cards)
What is the major difference between DNA replication on the leading and lagging strand?
Leading strand is continuous
Lagging strand is discontinuous
Define semi-conservative in the context of DNA synthesis
each strand of double stranded DNA serves as template for synthesis of new strand -> when 2 new molecules of DNA are formed each has 1 NEW & 1 OLD STRAND
Define bidirectional in the context of DNA synthesis
Replication is seen at both sides of replication fork (always 5’ to 3’ but goes in opposite directions- relatively- on leading and lagging strands)
Describe Okazaki fragments
fragments of discontinuous lagging strand synthesized in 5’ to 3’ direction, OPPOSITE OF FORK MOVEMENT
- few hundred to few thousand bp long
- synthesis stops when it gets to RNA primer
Describe the origin in the context of DNA synthesis
unique point where replication loops always begin, in circular DNA the 2 replication forks ultimately meet opposite origin
Describe the replication fork
Point where parent DNA is being unwound and separated strands are quickly replicated
What is the function of the origin binding protein?
Recognize the replication origin
What is the function of a helicase?
Melt/ unwind DNA
What is the function of single stranded binding proteins (SSB)?
Protect/ stabilize unwound DNA strands (single stranded DNA)
What is the function of primase?
responsible for synthesis of RNA primer, primosome constituent
What is the function of DNA polymerase I?
Fills gaps
excises primers
removes RNA primer
Requires template & RNA primer
What is the function of DNA polymerase III?
Elongate DNA from RNA primer
What is the function of DNA ligase?
Ligates or connects DNA fragments
i. e. Okazaki fragments
- ATP as cofactor in Eukaryotes
- NAD+ as cofactor in prokaryotes
What is the function of the sliding clamp?
Pol I is inefficient in synthesis because it goes on and off
Pol II associates with sliding protein that clamps on to DNA strand and prevents disassociation
Increases efficiency of synthesis
Increases processivity of DNA polymerase
PCNA is sliding clamp in Eukaryotic DNA
Pol gamma-beta subunit is sliding clamp of prokaryotic DNA Pol III
What is the function of Topisomerase/ gyrase?
Relieve torsional strain in front of replication fork - breaking and rejoining double stranded DNA
Prevents over-winding
What is the function of telomerase?
adds telomeres to stabilize DNA
What is the function of reverse transcriptase?
RNA dependent DNA polymerase (DNA from RNA)
Describe how DNA polymerase creates phosphodiester bond during addition of deoxyribonucleotides (dNTPs)
DNA pol I requires 3’ -OH on growing strand to act as primer and attack phosphoanhydride bond on next nucleotide
New nucleotide is added here with 3’ -OH primer for next bp addition
Addition based on pairing with template (non-coding) strand
=> dNMP (DNA) + dNTP -> dNMP (lenthend DNA) + PPi
What is the function of exonuclease?
Proofread DNA polymerase and correct errors
Describe the order of events that occur during, differences between, and coordination of DNA synthesis on leading and lagging strands
Leading strand ->
- Continuous copying following RNA primer after addition of ORC, follows helicase and gyrase/topoisomerase
- Moves in same direction as replication fork
Lagging Strand ->
- Uses RNA primer from previous okazaki fragment -> synthesizes 5’ to 3’ -> segments of varying lengths -> connected by ligases
- Discontinuous copying
- End replication problem – lagging strand cannot be synthesized to end during DNA replication (telomeres continuously shorten)
- Need RNA primer to begin synthesis of each piece of lagging strand DNA- at end there is no place for this to attach
- => ultimately telomeres are short enough that they initiate signal for cell death (apoptosis)
- In cancer telomerase enzyme de-repressed – restores telomeres – blocks normal cell death of old cell
Why is DNA repair so important?
- Damaged RNA quickly digested-> low impact
- DNA is irreplaceable => imperative to maintain
- Change in DNA=> mutation (permanent change to bp sequence) Mutation
- Often results in cancer
- DNA is only repaired macromolecule
How does DNA damage occur?
During replication From products of normal cell metabolism Radiation Chemicals (alkylation) Loss (depurination, depyrimidation, deamination) UV exposure
What are the 3 types of mutation?
Point mutation
Insertion
Deletion
What specific type of DNA damage does UV exposure cause?
Thymidine dimer