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SUMMARY:In vitro and in vivo single molecule studies of chromosome replica
 tion and segregation - David Sherratt\, University of Oxford
DTSTART:20110207T161500Z
DTEND:20110207T180000Z
UID:TALK29638@talks.cam.ac.uk
CONTACT:Scientific Meetings Co-ordinator
DESCRIPTION:A challenge of modern biology is to be able to observe exactly
  where and when molecular machines assemble and act within living cells. T
 his requires techniques that avoid ensemble averaging. By using slimfield 
 fluorescence microscopy\, we can observe single protein molecules with a 3
  ms temporal resolution and 5 nm spatial precision within live E. coli. Th
 is has allowed us to determine the in vivo stoichiometry and architecture 
 of the replisome and other molecular machines\, for example MukBEF that ac
 ts in chromosome organization. Sister replisomes track independently along
  the DNA and do not act as part of a replication factory [Cell 133\, 90\, 
 2008]. Each replisome contains 3 molecules of the replicative polymerase a
 nd the sliding clamp\, and single replicative helicases and clamp loaders 
 [Science 328\, 498\, 2010]. In addition to being able to observe the molec
 ular machines in vivo\, we are exploiting techniques that allow us to pert
 urb these machines by using controlled rapid degradation of specific prote
 ins. In complementary experiments\, we exploit in vitro single molecule st
 udies using ‘magnetic tweezers’ and ‘DNA curtains’ to gain insight
  into the function of proteins that act in chromosome segregation [EMBO J 
 29\, 1423\, 2010].
LOCATION:Max Perutz Lecture Theatre\, Medical Research Council (MRC) (MRC 
 Laboratory of Molecular Biol
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