Title | Measurement of the copy number of the master quorum-sensing regulator of a bacterial cell. |
Publication Type | Journal Article |
Year of Publication | 2010 |
Authors | Teng, S-W, Wang, Y, Tu, KC, Long, T, Mehta, P, Wingreen, NS, Bassler, BL, Ong, NP |
Journal | Biophys J |
Volume | 98 |
Issue | 9 |
Pagination | 2024-31 |
Date Published | 2010 May 19 |
ISSN | 1542-0086 |
Keywords | Gene Dosage, Microscopy, Fluorescence, Quorum Sensing, Repressor Proteins, Time Factors, Trans-Activators, Vibrio |
Abstract | <p>Quorum-sensing is the mechanism by which bacteria communicate and synchronize group behaviors. Quantitative information on parameters such as the copy number of particular quorum-sensing proteins should contribute strongly to understanding how the quorum-sensing network functions. Here, we show that the copy number of the master regulator protein LuxR in Vibrio harveyi can be determined in vivo by exploiting small-number fluctuations of the protein distribution when cells undergo division. When a cell divides, both its volume and LuxR protein copy number, N, are partitioned with slight asymmetries. We measured the distribution functions describing the partitioning of the protein fluorescence and the cell volume. The fluorescence distribution is found to narrow systematically as the LuxR population increases, whereas the volume partitioning is unchanged. Analyzing these changes statistically, we determined that N = 80-135 dimers at low cell density and 575 dimers at high cell density. In addition, we measured the static distribution of LuxR over a large (3000) clonal population. Combining the static and time-lapse experiments, we determine the magnitude of the Fano factor of the distribution. This technique has broad applicability as a general in vivo technique for measuring protein copy number and burst size.</p> |
DOI | 10.1016/j.bpj.2010.01.031 |
Alternate Journal | Biophys J |
PubMed ID | 20441767 |
PubMed Central ID | PMC2862190 |
Grant List | R01 GM065859 / GM / NIGMS NIH HHS / United States P50GM071508 / GM / NIGMS NIH HHS / United States 5R01AI054442 / AI / NIAID NIH HHS / United States / HHMI / Howard Hughes Medical Institute / United States K25 GM086909-02 / GM / NIGMS NIH HHS / United States R01 AI054442 / AI / NIAID NIH HHS / United States K25 GM086909 / GM / NIGMS NIH HHS / United States R01 GM065859-08 / GM / NIGMS NIH HHS / United States 5R01GM065859 / GM / NIGMS NIH HHS / United States P50 GM071508 / GM / NIGMS NIH HHS / United States R01 GM082938 / GM / NIGMS NIH HHS / United States |