Quantitative Characterization of Combinatorial Transcriptional Control of the Lactose Operon of E. coli

Biology – Quantitative Biology – Molecular Networks

Scientific paper

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Scientific paper

10.1073/pnas.0606717104

It is the goal of systems biology to understand the behavior of the whole in terms of the knowledge of the parts. This is hard to achieve in many cases due to the difficulty of characterizing the many constituents and their complex web of interactions involved in a biological system. The lac promoter of E. coli offers a possibility of confronting system-leve properties of transcriptional regulation with the known biochemistry of the molecular constituents and their mutual interactions. Such confrontations can reveal previously unknown constituents and interactions, as well as offering new insight into how the components work together as a whole. Here we study the combinatorial control of the lac promoter by the regulators LacR and CRP. A previous in vivo study [Setty et al., PNAS 100: 7702-7 (2003)] found gross disagreement between the observed promoter activites and the expected behavior based on the known molecular mechanisms. We repeated the study by identifying and removing several extraneous factors which significantly modulated the expression of the lac promoter. Through quantitative, systematic characterization of promoter activity for a number of key mutants and guided by the thermodynamic model of transcriptional gene regulation, we are able to account for the combinatorial control of the lac promoter quantitatively, in terms of a cooperative interaction between CRP and LacR-mediated DNA looping. Specifically, our analysis indicates that the sensitivity of the inducer response results from LacR-mediated DNA looping, which is significantly enhanced by CRP.

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