The cell cycle of Halobacterium salinarum: proteome changes and protein dynamics

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The cell cycle comprises events that must occur in all newborn cells before they are competent to divide again. It includes mass increase by a factor of two, exact doubling of the genome, segregation of the chromosomes, and cell division. It has intensively been studied with eukaryotes for decades. In recent years, it was revealed that also in bacteria cell cycle events are very tightly regulated in space and time. Much less is known about the cell cycle and its control in archaea. Halobacterium salinarum can be synchronized and thus it is ideally suited for cell cycle characterization. A previous study has revealed that only 79 genes are regulated on the transcript level and indicated that posttranscriptional regulatory mechanisms predominate in H. salinarum. Therefore, this project will concentrate on protein level regulation. Proteome analyses will identify proteins with cell cycle-dependent oscillations in their concentration as well as their post-translational modification, with a specific focus on differential protein phospharylation. GFP fusions with selected cell cycle proteins will reveal their specific intracellular localization and dynamic cell cycle-dependent changes. Conditional gene silencing will be used to study the molecular biological function of selected essential cell cycle proteins. Taken together, the results will yield an unprecedented insight into an archaeal cell cycle and its regulation.