@article{1747, keywords = {Trans-Activators, Recombinant Fusion Proteins, Gene Expression Regulation, Bacterial, Vibrio, Bacterial Proteins, Quorum Sensing, Repressor Proteins, Kinetics, Fluorescence, Colony Count, Microbial, Luminescent Proteins}, author = {Yufang Wang and Kimberly Tu and N Ong and Bonnie Bassler and Ned Wingreen}, title = {Protein-level fluctuation correlation at the microcolony level and its application to the Vibrio harveyi quorum-sensing circuit.}, abstract = {
Gene expression is stochastic, and noise that arises from the stochastic nature of biochemical reactions propagates through active regulatory links. Thus, correlations in gene-expression noise can provide information about regulatory links. We present what to our knowledge is a new approach to measure and interpret such correlated fluctuations at the level of single microcolonies, which derive from single cells. We demonstrated this approach mathematically using stochastic modeling, and applied it to experimental time-lapse fluorescence microscopy data. Specifically, we investigated the relationships among LuxO, LuxR, and the small regulatory RNA qrr4 in the model quorum-sensing bacterium Vibrio harveyi. Our results show that LuxR positively regulates the qrr4 promoter. Under our conditions, we find that qrr regulation weakly depends on total LuxO levels and that LuxO autorepression is saturated. We also find evidence that the fluctuations in LuxO levels are dominated by intrinsic noise. We furthermore propose LuxO and LuxR interact at all autoinducer levels via an unknown mechanism. Of importance, our new method of evaluating correlations at the microcolony level is unaffected by partition noise at cell division. Moreover, the method is first-order accurate and requires less effort for data analysis than single-cell-based approaches. This new correlation approach can be applied to other systems to aid analysis of gene regulatory circuits.
}, year = {2011}, journal = {Biophys J}, volume = {100}, pages = {3045-53}, month = {06/2011}, language = {eng}, }