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Title: Phosphotyrosine-mediated LAT assembly on membranes drives kinetic bifurcation in recruitment dynamics of the Ras activator SOS

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [2];  [1];  [1];  [1];  [1];  [1];  [3];  [1]
  1. Univ. of California, Berkeley, CA (United States). Dept. of Chemistry
  2. Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst.; Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
  3. Univ. of California, Berkeley, CA (United States). Dept. of Chemistry; Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst.; Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
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The assembly of cell surface receptors with downstream signaling molecules is a commonly occurring theme in multiple signaling systems. However, little is known about how these assemblies modulate reaction kinetics and the ultimate propagation of signals. Here, we reconstitute phosphotyrosine-mediated assembly of extended linker for the activation of T cells (LAT):growth factor receptor-bound protein 2 (Grb2):Son of Sevenless (SOS) networks, derived from the T-cell receptor signaling system, on supported membranes. Single-molecule dwell time distributions reveal two, well-differentiated kinetic species for both Grb2 and SOS on the LAT assemblies. The majority fraction of membrane-recruited Grb2 and SOS both exhibit fast kinetics and single exponential dwell time distributions, with average dwell times of hundreds of milliseconds. The minor fraction exhibits much slower kinetics, extending the dwell times to tens of seconds. Considering this result in the context of the multistep process by which the Ras GEF (guanine nucleotide exchange factor) activity of SOS is activated indicates that kinetic stabilization from the LAT assembly may be important. This kinetic proofreading effect would additionally serve as a stochastic noise filter by reducing the relative probability of spontaneous SOS activation in the absence of receptor triggering. In conclusion, the generality of receptor-mediated assembly suggests that such effects may play a role in multiple receptor proximal signaling processes.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE; National Institutes of Health (NIH)
Grant/Contract Number:
AC02-05CH11231; P01 AI091580
OSTI ID:
1379525
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, Issue 29; ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)Copyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 70 works
Citation information provided by
Web of Science

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Cited By (11)

Multicomponent Supported Membrane Microarray for Monitoring Spatially Resolved Cellular Signaling Reactions journal February 2018
Integrin and cadherin clusters: A robust way to organize adhesions for cell mechanics journal December 2016
Regulatory mechanisms in T cell receptor signalling journal May 2018
Understanding T cell signaling using membrane reconstitution journal August 2019
A molecular assembly phase transition and kinetic proofreading modulate Ras activation by SOS journal March 2019
Stoichiometry controls activity of phase-separated clusters of actin signaling proteins journal March 2019
Mapping the stochastic sequence of individual ligand-receptor binding events to cellular activation: T cells act on the rare events journal January 2019
More from less – bottom-up reconstitution of cell biology journal February 2019
Light-based tuning of ligand half-life supports kinetic proofreading model of T cell signaling journal April 2019
Kinetics of cytokine receptor trafficking determine signaling and functional selectivity journal November 2019
Multiple sources of signal amplification within the B cell Ras/MAPK pathway journal September 2018

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60 APPLIED LIFE SCIENCES
59 BASIC BIOLOGICAL SCIENCES
signal transduction
protein assembly
kinetic proofreading
membrane dwell time
single molecule