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Dr. GoldenWe apply a spectrum of approaches to achieve a comprehensive understanding of the circadian biological clock in Synechococcus elongatus. This cyanobacterium is the only developed model organism for studying a prokaryotic circadian system. We use biochemistry, bioinformatics, genetics, genomics, physiology, and structural biology to identify and determine the functions and interactions of components of the cyanobacterial clock. We have mutagenized approximately 90% of loci in the 2.7 Mb genome of S. elongatus and are surveying each mutant for defects in circadian rhythmicity in order to assemble a complete parts list for the clock and to identify the connections between the clock and fundamental cellular functions. Inactivation and overexpression of genes of interest, co-purification of their products with interacting partners, and structural determination of key components all help to elucidate the pathways that relay temporal information. Collaborations provide expertise in NMR, EPR, and mass spectrometry methods to provide insights into protein functions. Specific hypotheses query the mechanisms by which the clock becomes synchronized with the environment and the activities of redox-sensing components of that pathway. We are also assessing the contributions to timekeeping of subcellular localization and post-translational modifications of the three proteins that comprise the central oscillator of the cyanobacterial circadian clock. |
