Objectives * Characterisation of DNA binding by the antitoxin MazE. Three MazE dimers bind to three consecutive sites on a stretch of DNA of about 45 base pairs. The three individual sites A, B and C and the combinations AB and BC will be produced by annealing complementary synthetic oligonucleotides. Binding of MazE to these constructs will be measured to quantify the individual interactions and determine if there is cooperativity in the binding of the complete operator sequence. * Analysis of the compatibility between components from different systems (chromosomal versus plasmid-encoded; E. coli versus Vibrio) to gain insight in the evolution of the systems. This will also provide essential information for drug design purposes and the expected range of species that would be targetted by newly developed antibiotics. The chromosomal CcdBVf toxin from Vibrio fisheri has already been purified and appears to be very suitable for crystallographic and calorimetric studies. * Crystallisation of an antitoxin/DNA complex. The structure of a complex of an antitoxin with a relevant fragment of the operator DNA would provide insights into the regulation mechanisms of TA systems. The crystallisation of protein/DNA complexes has historically presented particular challenges, but a combination of stucture-based modelling with the aforementioned binding studies should provide sufficient information to identify a suitable DNA fragment.
|Effective start/end date||1/10/06 → 30/09/12|
Flemish discipline codes
- Biological sciences