Objective All living cells are surrounded by a membrane which provides a barrier between the contents of the cell and its environment. Various crucial processes in cells, such as cell/organelle division, or endo-/exo-cytosis, require the fusion of membranes. In e ukaryotes, various proteins have been identified that are involved in different membrane fusion events. In bacteria, membrane fusion occurs during cell division and sporulation. Recently, the first bacterial protein has been identified that is involved i n membrane fusion. This protein, SpolllE, is necessary for the membrane fusion reaction that occurs at a late stage in sporulation in Bacillus subtilis. SpolllE is a member of the SpolllE/FtsK family of proteins which is widely conserved in bacteria. Sp olllE/FtsK are involved in cell division and sporulation, and localise at the leading edge of the invaginating membrane. The proteins consist of an N-terminal region containing four transmembrane regions, a linker region, and a C-terminal domain that me diates energy-dependent DNA transport. During cell division, SpolllE/FtsK are critical for correct chromosome partitioning, whereas in sporulating bacteria, SpolllE transports DNA from the mother-cell into the prespore. This project focuses on the role of the N-terminal membrane segment of SpolllE/FtsK in membrane fusion and DNA transport. The following questions will be addressed: 1) Do the transmembrane regions of SpolllE/FtsK oligomerise to form a DNA pore? 2) Do the transmembrane regions of Spolll E/FtsK mediate membrane fusion? 3) What is the energy requirement of DNA transport across a biological membrane? Full-length SpolllE/FtsK and truncates containing only the transmembrane region will be purified and reconstituted in artificial liposome sys tems. These systems allow a detailed characterisation of both the membrane fusion and DNA transport reactions. Fields of science natural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesgeneticschromosomes Keywords DNA transport cell division membrane fusion sporulation Programme(s) FP6-MOBILITY - Human resources and Mobility in the specific programme for research, technological development and demonstration "Structuring the European Research Area" under the Sixth Framework Programme 2002-2006 Topic(s) MOBILITY-4.1 - Marie Curie European Reintegration Grants (ERG) Call for proposal FP6-2002-MOBILITY-11 See other projects for this call Funding Scheme ERG - Marie Curie actions-European Re-integration Grants Coordinator VRIJE UNIVERSITEIT AMSTERDAM Address De boelelaan 1105 Amsterdam Netherlands See on map EU contribution € 0,00
