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DNA Replication. Mechanism of DNA translocation in a replicative helicase. http://library.tedankara.k12.tr/chemistry/vol1/biochem/trans98.jpg. Helicase Function. Unwind dsDNA to make ssDNA DNA replication Repair Recombination.

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dna replication

DNA Replication

Mechanism of DNA translocation in a replicative helicase

http://library.tedankara.k12.tr/chemistry/vol1/biochem/trans98.jpg

helicase function
Helicase Function
  • Unwind dsDNA to make ssDNA
    • DNA replication
    • Repair
    • Recombination

http://library.tedankara.k12.tr/chemistry/vol1/biochem/trans98.jpg

helicase function1
Helicase Function
  • E1 & TAG (virus)
    • Recognize & melts origin to create ssDNA
    • Assemble on ssDNA
    • E1 3’-5’ to Unwind

http://library.tedankara.k12.tr/chemistry/vol1/biochem/trans98.jpg

slide4

Crystal structure of hexameric helicase (E1 of bovine papilomavirus)

  • Shows the path of DNA
  • Indicates mechanism
crystalization conditions
Crystalization Conditions
  • Fragment BPV-1 E1
    • Lacks N-term (306-577)
    • GST fusion
      • Concentrated with
        • 5 mM ADP
        • 200 mM MgCl2
        • Oligo T (13 mer)
    • Mother liquor 100 mM KCl
    • Crystals
      • Flash frozen (N2 (lq))
      • Synchotron Radiation
    • Molecular replacement

http://en.wikipedia.org/wiki/Image:X_ray_diffraction.png

slide7

5’

3’

loops
Loops

roll

staircase model
Staircase Model
  • A helicase staircase.Raney KD. Nat Struct Mol Biol. 2006 Aug;13(8):671-2.
slide20

ADP

Y534

ATP

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