Despite the prediction that ORC and DnaA function through a common mechanism, to date there has been no observed DNA unwinding activity by a eukaryotic initiator. To explore ORC's interaction with DNA, DmORC was imaged with various linear and supercoiled DNA substrates using AFM. These data suggest ORC is wrapping an average of a¼130bp of linear DNA. This study represents the first structural characterization of a metazoan initiator and allows for a more detailed comparison of the initiators across the three domains of life.The final reconstruction includes ~4, 500 particles and has a resolution of 34A as estimated by the Fourier Shell correlation (FSC) cut-off at 0.5 ... For the ATP-ORC structure, ATP-yS was added to glycerol gradient purified ORC to a ... Docking of the X-ray crystal structure of the ATP-DnaA model was done manually in PyMol.
|Title||:||Single Particle Structural Analysis of the Drosophila Melanogaster Origin Recognition Complex|
|Author||:||Megan Gretchen Clarey|
|Publisher||:||ProQuest - 2008|