DNA REPLICATION Flashcards
DNA STRUCTURE
Nucleic acids: Biopolymers built from smaller molecules (monomers) connected covalently.
DNA structure: Double helix with a sugar-phosphate backbone and nitrogenous bases (A, G, C, T).
DNA replication overview
Goal: To ensure each daughter cell receives one copy of DNA.
Semi-conservative replication: Each new DNA molecule has one old and one new strand.
Other theories: Conservative and dispersive models (disproven).
challenges
- Replicate all bases of long DNA accurately.
- Minimize replication errors.
- Overcome mechanical stress during unwinding and encounters with transcribed genes.
DNA polymerase and proofreading
DNA polymerase: Efficient at proofreading, with 1 error per 10^7 bases.
Exonuclease activity: Removes mismatches during replication to ensure accuracy.
basic nucleotide structure
DNA: A, G, C, T bases with deoxyribose sugar.
RNA: A, G, C, U bases with ribose sugar.
Sugar-phosphate backbone supports 5’ to 3’ polymerization.
bacterial genome replication
Circular DNA: Replication begins at a single origin (oriC) and proceeds bidirectionally.
Replication is not synchronized with cell division.
Plasmids: May have variable copy numbers.
initiation of DNA replication
oriC: A-T rich region where DNA melts due to weaker bonds.
DnaA protein binds to the origin, causing strand separation and allowing helicase attachment.
supercooling and topoisomerase
As DNA unwinds, supercoiling occurs.
Topoisomerases: Remove supercoiling to prevent replication machinery from stalling.
leading and lagging strands
Leading strand: Continuous synthesis in the 5’ to 3’ direction.
Lagging strand: Synthesized in short fragments (Okazaki fragments) joined by DNA ligase.
key replication enzymes
DNA polymerase III: Extends the DNA strand but cannot start it.
Primase: Synthesizes an RNA primer for DNA pol III to extend.
Helicase: Unwinds the DNA strands.
DNA polymerase I: Removes the RNA primer and fills gaps on the lagging strand.
DNA ligase: Joins Okazaki fragments.
sugar structure in nucleotides
C1: Binds the base.
C2: No OH in DNA; OH present in RNA.
C3 and C5: Interact with phosphate groups for polymerization.
double stranded DNA rules
Anti-parallel strands.
Base-pairing: A with T, G with C.
Nucleotides added to the 3’ end using energy from phosphate bond hydrolysis.
polarity in DNA
5’ end: Phosphate group.
3’ end: Hydroxyl group.
Results in directionality from 5’ to 3’ in synthesis.
eukaryotic genome replication
Requires multiple origins due to larger, linear chromosomes.
Euchromatin origins fire first (early origins).
Heterochromatin origins fire later (late origins).
Replication occurs during the S-phase of the cell cycle.
replicons
DNA region replication from a single origin
Bacteria: Each circular DNA is a single replicon.
Eukaryotes: Have multiple replicons, and not all origins fire in every cycle. Some are dormant.