Two transcriptional regulators of Pyrococcus have been characterized preliminarily in this laboratory. The Leucine responsive regulatory protein (LRP) and a 24 kDa regulator of a heat shock regulon. The interaction of these components with the basal transcriptional machinery and their mechanism of action will be studied in molecular detail. DNA-protein interactions will be investigated by various footprinting techniques, protein-protein interactions will be studied in vitro and in vivo. The crystal structure of the heat shock regulator both alone and in complex with its target DNA will be determined. The effect of NTP concentration on transcription of various promoters directing expression of proteins involved in pyrimidine synthesis will be analysed. *
