Dna replication
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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

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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


Dna replication

  • 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


Dna replication

5’

3’


Dna replication

Courtesy of Chayne Piscitelli


Loops

Loops

roll


D504 r505 k508

D504, R505 & K508


Staircase model

Staircase Model

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


Dna replication

ATP?


Dna replication

ADP

Y534

ATP


Escort model

Escort Model


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