Abstract
Abstract:In order for a cell to function normally in any environmental condition, a cell must be able to perform basic cellular functions such as cell metabolism, transcription, translation, and cell division. Adaptation to extremes of physicochemical conditions such as high pressure and temperature requires that all basic cellular processes are unrestricted. Here, we focus on the effect of pressure and temperature on transcription. For the initiation of transcription to occur, RNA polymerase must bind to DNA. We have studied the binding propensity of T7-RNA polymerase and Escherichia coli (E. coli) RNA polymerase to a consensus T7 promoter and E.coli recA promoter DNA, respectively. Specifically, we have investigated the stability of RNA polymerase-promoter DNA elongation complex in a wide range of pressure (1-2000 atm) and temperature (10-50oC) using fluorescence anisotropy. At constant temperature, the stability of elongation complex decreases with increasing pressure. The pressure at which transcription elongation stops depends on temperature. We construct a pressure-temperature phase diagram of the stability of elongation complex for both T7 polymerase + T7 promoter and E.coli polymerase + recA promoter. Furthermore, we show that the pressure-temperature stability phase diagram of elongation complex exhibits typical elliptical shape usually associated with the stability of proteins. Our results suggest that the evolution of polymerases has maintained their functionality in a wide range of pressures and temperatures.
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