DNA Flashcards
Stabilizing Forces of DNA, and Structure
- Comp bases are H bonded
- AT have 2 H binds
- GC have 3 H bonds
- Hydrophilic phosphate backbone
- Hydrophobic bases stacked inside helix
- Strands are coiled around a common axis of symmetry
- DS DNA has major and monor grooves
What causes denaturation
- DNA seperates when H bonds are disrupted
- Alteration of pH
- Increased heat
- DNA replication
- Complementary DNA strands can reanneal (hybridize)
- Tm is temp at which 1/2 DNA is denatured
- GC has a higher Tm than AT
Organization of DNA
46 choromosomes
DNA associated with histones forming Nucleosomes
Nucleosomes further arranged into increasingly comlex structures and eventually chromosomes
Gen steps in DNA synthesis
- ID origins of replication
- Unwinding (denaturation) of ds DNA to provide ss DNA template
- Formation of replication fork and synthesis of RNA primer
- Initiation of DNA synthesis, elongation, and ligation of newly synthesized DNA segments
- Reconstitution of chromatin structure
Where does DNA rep occur
Direction
Requires…
- Occurs at replication fork and is bidirectional
- Rep fork has leading and lagging (Okazaki) strands
- Semiconservative
- Proceeds 5 to 3
- Requires RNA primer
- High processivity
- Base selected based on H bonding and geometry
- High fidelity (low error rate)
DnaA
Replication initiation factor
Promotes denaturation of DNA at origin of replication
DNA Helicase
Binds ssDNA near the replication fork and uses ATP to unzip DNA
SS DNA binding protein
Keeps DNA ss
Topoisomerase
Cuts one or both strands of DNA to change DNA topology and relieve coiling
Primase
Synthesizes RNA primers to prime DNA sysnthesis
DNA polymerases
Replicate DNA
Ligase
Links DNA by catalyzing phosphodiester bonds
Occurs during DNA rep by joining Okazaki fragments
DNA repair
DNA Poly III
Adds correct nucleotide thru formation of phosphodiester bond
Exonuclease activity hydrolyzes phoshodiester bond
5–>3 activity, base removal from 5 terminus, removes RNA primer
3–>5 exonuclease activity mediates proofreading, Base removal from 3 terminus, removes misincorporated bases