Structural analysis of the genetic switch controlling expression of the AhdI restriction-modification system

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The regulation of gene expression in all organisms is dependent on complex interactions between gene-regulatory proteins and their DNA targets. A full understanding of such mechanisms requires analysis at the biophysical and biochemical level. Here, we investigate the molecular basis of the genetic switch involved in the regulation of bacterial restriction-modification systems. Restriction-modification systems in bacteria act as a form of primitive 'immune system'; they recognise foreign DNA sequences and are able to destroy invading DNA. However, expression of the gene for the enzyme that destroys the DNA must be delayed, to allow time for the host DNA to become labelled as 'self' (by a process called methylation) and thus protected from cleavage. A controller protein is responsible for this timing delay, and we have solved its molecular structure by X-ray diffraction. We have also established that the protein forms a number of complexes when it interacts with its target DNA sequence. The aim of this project is to discover the 3-dimensional structure of the various regulatory complexes bound to DNA, and to understand how they interact to allow regulation of the relevant genes. Together, these experimental approaches should provide a detailed molecular picture of the genetic switch underlying the establishment and maintenance of restriction-modification systems, many features of which should be generally applicable to a wide range of gene regulatory systems.