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A Fyn-specific biosensor reveals localized, pulsatile kinase activity and spatially regulated signaling crosstalk

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Title A Fyn-specific biosensor reveals localized, pulsatile kinase activity and spatially regulated signaling crosstalk
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Creator Ananya Mukherjee, Randhir Singh, Sreeram Udayan, Sayan Biswas, Purushotham Pothula, Saumya Manmadhan, Geen George, Shilpa Dilip Kumar, Ranabir Das, Balaji M Rao, Akash Gulyani
 
Subject Not Available
 
Description Not Available
Cell behavior is controlled through spatio-temporally localized protein activity. Despite unique and often contradictory roles played by Src-family-kinases (SFKs) in regulating cell physiology, activity patterns of individual SFKs have remained elusive. Here, we report a biosensor for specifically visualizing active conformation of SFK-Fyn in live cells. We deployed combinatorial library screening to isolate a binding-protein (F29) targeting activated Fyn. Nuclear-magnetic-resonance (NMR) analysis provides the structural basis of F29 specificity for Fyn over homologous SFKs. Using F29, we engineered a sensitive, minimally-perturbing fluorescence-resonance-energy-transfer (FRET) biosensor (FynSensor) that reveals cellular Fyn activity to be spatially localized, pulsatile and sensitive to adhesion/integrin signaling. Strikingly, growth factor stimulation further enhanced Fyn activity in pre-activated intracellular zones. However, inhibition of focal-adhesion-kinase activity not only attenuates Fyn activity, but abolishes growth-factor modulation. FynSensor imaging uncovers spatially-organized, sensitized signaling clusters, direct crosstalk between integrin and growth-factor-signaling, and clarifies how compartmentalized Src-kinase activity may drive cell fate.
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Date 2019-12-03T10:12:12Z
2019-12-03T10:12:12Z
2019-01-01
 
Type Research Paper
 
Identifier Not Available
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http://krishi.icar.gov.in/jspui/handle/123456789/26683
 
Language English
 
Relation Not Available;
 
Publisher Cold Spring Harbor Laboratory