A two-component protein condensate of the EGFR cytoplasmic tail and Grb2 regulates Ras activation by SOS at the membrane
Edited by Natalie Ahn, University of Colorado Boulder, Boulder, CO; received December 17, 2021; accepted March 22, 2022
Significance
Two-dimensional condensates of proteins on the membrane surface, driven by tyrosine phosphorylation, are beginning to emerge as important players in signal transduction. This work describes discovery of a protein condensation phase transition of EGFR and Grb2 on membrane surfaces, which is poised to have a significant impact on how we understand EGFR signaling and misregulation in disease. EGFR condensation is mediated through a Grb2-Grb2 crosslinking element, which itself is regulatable through a specific phosphotyrosine site on Grb2. Furthermore, the EGFR condensate exerts significant control over the ability of SOS to activate Ras, thus implicating the EGFR condensate as a regulator of signal propagation from EGFR to Ras and the MAPK pathway.
Abstract
We reconstitute a phosphotyrosine-mediated protein condensation phase transition of the ∼200 residue cytoplasmic tail of the epidermal growth factor receptor (EGFR) and the adaptor protein, Grb2, on a membrane surface. The phase transition depends on phosphorylation of the EGFR tail, which recruits Grb2, and crosslinking through a Grb2-Grb2 binding interface. The Grb2 Y160 residue plays a structurally critical role in the Grb2-Grb2 interaction, and phosphorylation or mutation of Y160 prevents EGFR:Grb2 condensation. By extending the reconstitution experiment to include the guanine nucleotide exchange factor, SOS, and its substrate Ras, we further find that the condensation state of the EGFR tail controls the ability of SOS, recruited via Grb2, to activate Ras. These results identify an EGFR:Grb2 protein condensation phase transition as a regulator of signal propagation from EGFR to the MAPK pathway.
Data Availability
All study data are included in the article and/or SI Appendix.
Acknowledgments
We thank the members of the Kuriyan Laboratory and the Groves Laboratory for helpful discussion and critical feedback. This work was supported by NIH National Cancer Institute Physical Sciences in Oncology Network Project 1-U01CA202241 and by the Novo Nordisk Foundation Challenge Program under the Center for Geometrically Engineered Cellular Systems. Additional support was provided by NIH Grant PO1 A1091580.
Supporting Information
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Cover image: Pictured is a solar flare imaged by NASA's Solar Dynamics Observatory on October 2, 2014. Mang Lin and Mark H. Thiemens measured sulfur-35 isotopes, produced via cosmic-ray interaction with atmospheric argon, weekly between 2014 and 2016 in La Jolla, California. Sulfur-35 concentrations during the study period, which included a solar activity maximum, was lower than measurements made during the 2009–2010 solar activity minimum, suggesting a link between sulfur-35 production and solar activity. The authors also found links between sulfur-35 concentrations and Elñio events, suggesting that sulfur-35 may help uncover solar, weather, and climate effects on atmospheric radioisotope production and transport. See the article by Lin and Thiemens, e2121550119. Image credit: NASA Goddard Space Flight Center.
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Copyright
Copyright © 2022 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).
Data Availability
All study data are included in the article and/or SI Appendix.
Submission history
Received: December 17, 2021
Accepted: March 22, 2022
Published online: May 4, 2022
Published in issue: May 10, 2022
Keywords
Acknowledgments
We thank the members of the Kuriyan Laboratory and the Groves Laboratory for helpful discussion and critical feedback. This work was supported by NIH National Cancer Institute Physical Sciences in Oncology Network Project 1-U01CA202241 and by the Novo Nordisk Foundation Challenge Program under the Center for Geometrically Engineered Cellular Systems. Additional support was provided by NIH Grant PO1 A1091580.
Notes
This article is a PNAS Direct Submission.
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Competing Interests
The authors declare no competing interest.
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