L7: DNA Replication Flashcards
Learning Objectives: • Outline the mechanism of DNA replication in prokaryotes. • Outline the importance and processes of proofreading and error correction by DNA polymerases. Outline key differences between the mechanisms of DNA replication in prokaryotes and eukaryotes.
Why is DNA replication important?
New cells are created by mitosis, which required DNA replication
What happens in DNA replication?
Identical copy of the parent DNA is used as a template for a new strand
Define semi-conservative replication
2 copies of the original DNA is produced, consisting of only half of the material
When is the replication fork created?
DNA helicase unwinds the double helix structure
Initiation of DNA replication in prokaryotes
1) Initiation proteins bind to replication origin
2) Separate strands of DNA
3) Two ‘replication forks’ are formed
Which directions do replication forks move?
In opposite directions
Which bonds are nucleotides joined by?
Phosphodiester Bonds
Structure of deoxynucleoside triphosphates (dNTPs)
3 phosphate groups attached to the 5’ carbon of deoxyribose
Role of DNA polymerase III
Synthesis the bulk of DNA in E.coli
What is a holoenzyme?
Composed of multiple subunits with different functions
What does DNA pol III require for DNA synthesis?
RNA primer
How is RNA primer created ?
Using enzyme PRIMASE
Which direction does DNA pol III synthesis new DNA in?
5’ to 3’ direction
What is the leading strand?
Replicates continuously
What is the lagging strand?
Replicates to form short fragments
Polarity problem in DNA replication
DNA double helix have opposite polarity but all DNA polymerase replicate in the same direction
What are Okazaki fragments joined by?
DNA ligase
Role of ligase in DNA replication
Joins Okazaki fragments into continuous DNA strands
Function of enzyme topoisomerase
Preventing DNA from getting tangled
- Relieving pressure supercoiling ahead of replication fork
Role of Type I topoisomerases
Cut 1 DNA strand
Role of Type II topoisomerases
Cut both DNA strands, pass coil through gaps (ATP) & reseal the strands
Role of helicase in DNA replication
Separates DNA strands
Role of single-stranded binding protein in DNA replication
Prevents DNA strands from re-annealing
Role of primase in DNA replication
Synthesise RNA primers