期刊
CELL
卷 157, 期 7, 页码 1724-1734出版社
CELL PRESS
DOI: 10.1016/j.cell.2014.04.039
关键词
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资金
- Human Frontier Science Program (HFSP) postdoctoral fellowship [LT000529/2012-L]
- NIH [5R21AI104305-02, 5DP1LM01150-05]
- Stanford Center for Systems Biology [NIH P50GM107615]
- Allen Distinguished Investigator Award
Increasing evidence has shown that population dynamics are qualitatively different from single-cell behaviors. Reporters to probe dynamic, single-cell behaviors are desirable yet relatively scarce. Here, we describe an easy-to-implement and generalizable technology to generate reporters of kinase activity for individual cells. Our technology converts phosphorylation into a nucleocytoplasmic shuttling event that can be measured by epifluorescence microscopy. Our reporters reproduce kinase activity for multiple types of kinases and allow for calculation of active kinase concentrations via a mathematical model. Using this technology, we made several experimental observations that had previously been technically-unfeasible, including stimulus-dependent patterns of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NF-kappa B) activation. We also measured JNK, p38, and ERK activities simultaneously, finding that p38 regulates the peak number, but not the intensity, of ERK fluctuations. Our approach opens the possibility of analyzing a wide range of kinase-mediated processes in individual cells.
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