Final Report Summary - INTERCOM (The Influence of Interfaces, Confinement and Compartmentalization on Chemical Reactions) The cell is the basic unit of life. The interior of living cells is a unique, strongly crowded environment and there are significant gaps in our knowledge and understanding of how crowding impacts on complex enzymatic processes and non-covalent interactions between cellular building blocks. In other words, the diffusion and activity as well as interactions between proteins, DNA and RNA are all very different inside the cellular environment. Indeed the crowded conditions in the cell strongly favor formation of macromolecular complexes and it is surprising that weak non-covalent interactions do not completely lead to a ‘freezing’ of all macromolecules within cell. In our research, we have discovered how coacervation of collections of proteins and other molecules can form compartments capable of transcription and translation (i.e. the process of producing proteins encoded by DNA). Coacervation creates an artificial cell-like environment in which the binding constant between DNA and RNA-polymerase is larger by two orders of magnitude and the rate of mRNA production is increased by a factor of four. The effect of crowding on the kinetics of the fundamental machinery of gene expression has a direct impact on our understanding of biochemical networks in vivo and will aid the development of synthetic cells.
