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SUMMARY:Proteomic and cell biological approaches to the study of clathrin 
 in a vertebrate conditional knockout cell-line - Dr. Tony Jackson\, Univ. 
 Cambridge\, Neuroscience
DTSTART:20101119T140000Z
DTEND:20101119T150000Z
UID:TALK26615@talks.cam.ac.uk
CONTACT:Erika Eiser
DESCRIPTION:In eukaryotic cells\, clathrin is required for the selective t
 ransport of proteins between intracellular membrane compartments. To under
 stand the role of clathrin in vivo\, we exploited the unique properties of
  the chicken B cell-line DT40. This cell-line has become established as an
  important tool in cell biology because it exhibits a remarkably high rate
  of homologous recombination\, thus allowing targeted gene knockouts to be
  easily generated. We replaced endogenous clathrin genes in DT40 with a cl
 athrin gene under the control of a regulatable promoter. Hence cells can b
 e grown either with clathrin or without and their properties compared. Usi
 ng a whole cell proteomic methodology called LOPIT (Localisation of Organe
 lle Proteins by Isotope Tagging) we have completed the first proteomic ana
 lysis of the major organelles in DT40 and compared the changes that occur 
 when clathrin expression is repressed. In this talk I will explain how the
  LOPIT methodology works and will show how the use of DT40 has produced ne
 w insights into clathrin function.
LOCATION:Pippard Lecture Theatre\, Cavendish Laboratory
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