Path of DNA within Mu transpososomes: order, dynamics and topology of Mu end-enhancer interactions

dc.contributor.advisorJayaram, Makkunien
dc.contributor.advisorHarshey, Rasika M.en
dc.creatorPathania, Shailjaen
dc.description.abstractUsing a new method called ‘difference topology’, a complete topological analysis of all the intermediates in the Mu transposition pathway has been carried out. The results give us a first look into the intricate architecture of transpososomes made by the Mu transposase. We show that the three participating DNA sites - L (left end), R (right end) and E (enhancer) - are brought together to make an elaborate 5-noded synapse, wherein E crosses R twice and L once, and L and R cross each other two times. This is the most complex DNA arrangement seen to date within a recombination synapse. The order and dynamics of association of these sites was monitored within a series of transpososomes prior to and during synapsis (LER), engagement (type 0), cleavage (type I) and strand transfer (type II) of Mu ends. In the process, we have discovered a new intermediate ER, which likely initiates the assembly pathway. We have found that the enhancer, previously believed to have exited the transpososome as early as in the type 0 complex, instead remains within the transpososome through completion of transposition. Our results suggest that the enhancer may play additional roles in transposition than previously thought.
dc.rightsCopyright is held by the author. Presentation of this material on the Libraries' web site by University Libraries, The University of Texas at Austin was made possible under a limited license grant from the author who has retained all copyrights in the works.en
dc.subject.lcshBacteriophage muen
dc.titlePath of DNA within Mu transpososomes: order, dynamics and topology of Mu end-enhancer interactionsen