8. DNA Replication Flashcards
What are the requirements for DNA replication?
Single-stranded template Deoxyribonucleotide triphosphates Mg++ Replisome (protein complex) Primer with free 3' end hydroxyl group
What sequence is indicative of an origin of replication?
Short AT rich region
What is the function of DNA helicase?
Separate the DNA strands
Uses ATP
What is the function of single-strand binding proteins?
Prevent the strands from re-associating
What is the function of topoisomerase?
Regulates twisting of DNA
Can cut DNA
What is the function of primase (RNA polymerase enzyme)?
Makes a primer (RNA starter strand)
After replication, RNA primer is removed and the gap is filled by DNA polymerase
What is the function of DNA polymerase?
Uses single strand DNA as a template
Reads template 3’ to 5’
Makes new DNA from 5’ to 3’
Aligns and adds nucleotides along template which specifies the sequence of the new chain
Catalyses formation of phosphodiester bonds
What is the role of PCNA?
Acts as a sliding stamp to keep DNA Polymerase closely associated to the template
How are errors prevented?
Substrate specificity: Catalysis only occurs when correct dNTP is bound
‘Proof reading’: 3’ to 5’ exonuclease activity
Describe lagging strand synthesis
Primase makes a new primer at regular intervals
DNA polymerase replicates the template from the primer producing a new strand in the 5’ to 3’ direction
DNA Polymerase is blocked by the proximity to the next primer
Primers removed, gaps filled and backbone joined
What is the role of Polymerase alpha?
Initiating replication
Associates with primase
Replicates DNA by extending primer 5’ to 3’
What is the role of polymerase epsilon?
Replicates leading strand DNA by extending primer 5’ to 3’
3’ to 5’ exonuclease activity
What is the role of polymerase delta?
Replicates lagging strand DNA by extending primer 5’ to 3’
3’ to 5’ exonuclease activity
What is the role of polymerase beta?
Involved in DNA repair
What is the role of polymerase gamma?
Replicates mitochondrial DNA