CaltechAUTHORS
  A Caltech Library Service

Eph-ephrin signaling modulated by polymerization and condensation of receptors

Ojosnegros, Samuel and Cutrale, Francesco and Rodríguez, Daniel and Otterstrom, Jason J. and Chiu, Chi Li and Hortigüela, Verónica and Tarantino, Carolina and Seriola, Anna and Mieruszynski, Stephen and Martínez, Elena and Lakadamyali, Melike and Raya, Angel and Fraser, Scott E. (2017) Eph-ephrin signaling modulated by polymerization and condensation of receptors. Proceedings of the National Academy of Sciences of the United States of America, 114 (50). pp. 13188-13193. ISSN 0027-8424. https://resolver.caltech.edu/CaltechAUTHORS:20171130-140826020

[img] PDF - Published Version
See Usage Policy.

1630Kb
[img] PDF - Supplemental Material
See Usage Policy.

1131Kb
[img] PDF (Appendix) - Supplemental Material
See Usage Policy.

10Mb
[img] Video (MPEG) (Movie S1) - Supplemental Material
See Usage Policy.

605Kb
[img] Video (MPEG) (Movie S2) - Supplemental Material
See Usage Policy.

1303Kb
[img] Video (MPEG) (Movie S3) - Supplemental Material
See Usage Policy.

2358Kb
[img] Video (MPEG) (Movie S4) - Supplemental Material
See Usage Policy.

2618Kb
[img] Video (MPEG) (Movie S5) - Supplemental Material
See Usage Policy.

2184Kb
[img] Video (MPEG) (Movie S6) - Supplemental Material
See Usage Policy.

4Mb
[img] Video (MPEG) (Movie S7) - Supplemental Material
See Usage Policy.

7Mb

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20171130-140826020

Abstract

Eph receptor signaling plays key roles in vertebrate tissue boundary formation, axonal pathfinding, and stem cell regeneration by steering cells to positions defined by its ligand ephrin. Some of the key events in Eph-ephrin signaling are understood: ephrin binding triggers the clustering of the Eph receptor, fostering transphosphorylation and signal transduction into the cell. However, a quantitative and mechanistic understanding of how the signal is processed by the recipient cell into precise and proportional responses is largely lacking. Studying Eph activation kinetics requires spatiotemporal data on the number and distribution of receptor oligomers, which is beyond the quantitative power offered by prevalent imaging methods. Here we describe an enhanced fluorescence fluctuation imaging analysis, which employs statistical resampling to measure the Eph receptor aggregation distribution within each pixel of an image. By performing this analysis over time courses extending tens of minutes, the information-rich 4D space (x, y, oligomerization, time) results were coupled to straightforward biophysical models of protein aggregation. This analysis reveals that Eph clustering can be explained by the combined contribution of polymerization of receptors into clusters, followed by their condensation into far larger aggregates. The modeling reveals that these two competing oligomerization mechanisms play distinct roles: polymerization mediates the activation of the receptor by assembling monomers into 6- to 8-mer oligomers; condensation of the preassembled oligomers into large clusters containing hundreds of monomers dampens the signaling. We propose that the polymerization–condensation dynamics creates mechanistic explanation for how cells properly respond to variable ligand concentrations and gradients.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1073/pnas.1713564114DOIArticle
http://www.pnas.org/content/114/50/13188PublisherArticle
http://www.pnas.org/content/suppl/2017/11/30/1713564114.DCSupplementalPublisherSupporting Information
ORCID:
AuthorORCID
Fraser, Scott E.0000-0002-5377-0223
Additional Information:© 2017 National Academy of Sciences. Published under the PNAS license. Edited by Harry B. Gray, California Institute of Technology, Pasadena, CA, and approved October 31, 2017 (received for review August 1, 2017). Published online before print November 30, 2017. The authors thank Giulia Ossato, William Dempsey, and Nicolas Plachta for useful discussions, and acknowledge the Nikon Center of Excellence at ICFO-The Institute of Photonic Sciences. S.O. was supported by Marie Curie International Outgoing Fellowship 276282 within the EU Seventh Framework Programme FP7/2007-2013 and Postdoctoral Fellowships LT000109/2011 from the Human Frontier Science Program Organization and EX2009-1136 from the Ministerio de Educación, Programa Nacional de Movilidad de Recursos Humanos del Plan Nacional de I-D+i 2008-2011. F.C. was supported by grants from the Moore Foundation and NIH (R01 HD075605 and R01 OD019037). J.J.O. acknowledges financial support from ICFONEST+, funded by the Marie Curie COFUND (FP7-PEOPLE-2010-COFUND) action of the European Commission and by the MINECO Severo Ochoa action at ICFO-The Institute of Photonic Sciences. Additional funding was provided by the Generalitat de Catalunya (2014-SGR-1442 and 2014-SGR-1460); the Spanish Ministry of Economy and Competitiveness (SAF2015-69706-R, MINAHE5, TEC2014-51940-C2-2-R, SEV-2015-0522); Instituto de Salud Carlos III (ISCIII)/FEDER (RD16/0011/0024); the European Union (GLAM project GA-634928; System’s Microscopy Network of Excellence consortium FP-7-HEALTH.2010.2.1.2.2); the European Research Council (337191-MOTORS and 647863-COMIET); the Fundació Privada Cellex; and CERCA Programme/Generalitat de Catalunya. Ethics Statement: The experiments presented in this study were conducted following protocols approved by the Institutional Review Board of the Center of Regenerative Medicine in Barcelona. Author contributions: S.O., F.C., D.R., C.L.C., V.H., E.M., M.L., A.R., and S.E.F. designed research; S.O., F.C., D.R., J.J.O., C.L.C., V.H., C.T., A.S., S.M., and S.E.F. performed research; E.M. and S.E.F. contributed new reagents/analytic tools; S.O., F.C., D.R., J.J.O., C.L.C., V.H., S.M., and E.M. analyzed data; and S.O., F.C., D.R., J.J.O., C.L.C., V.H., E.M., M.L., A.R., and S.E.F. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1713564114/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
European Research Council (ERC)276282
Human Frontier Science ProgramLT000109/2011
Ministerio de EducaciónEX2009-1136
Gordon and Betty Moore FoundationUNSPECIFIED
NIHR01 HD075605
NIHR01 OD019037
Marie Curie FellowshipUNSPECIFIED
European CommissionUNSPECIFIED
Ministerio de Economía, Industria y Competitividad (MINECO)UNSPECIFIED
Generalitat de Catalunya2014-SGR-1442
Generalitat de Catalunya2014-SGR-1460
Ministerio de Economía, Industria y Competitividad (MINECO)SAF2015-69706-R
Ministerio de Economía, Industria y Competitividad (MINECO)MINAHE5
Ministerio de Economía, Industria y Competitividad (MINECO)TEC2014-51940-C2-2-R
Ministerio de Economía, Industria y Competitividad (MINECO)SEV-2015-0522
Instituto de Salud Carlos IIIRD16/0011/0024
European UnionGA-634928
European Research Council (ERC)337191-MOTORS
European Research Council (ERC)647863-COMIET
Fundació Privada CellexUNSPECIFIED
Subject Keywords:Eph; ephrin; receptor tyrosine kinase; gradients; cell communication
Issue or Number:50
Record Number:CaltechAUTHORS:20171130-140826020
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171130-140826020
Official Citation:Samuel Ojosnegros, Francesco Cutrale, Daniel Rodríguez, Jason J. Otterstrom, Chi Li Chiu, Verónica Hortigüela, Carolina Tarantino, Anna Seriola, Stephen Mieruszynski, Elena Martínez, Melike Lakadamyali, Angel Raya, and Scott E. Fraser Eph-ephrin signaling modulated by polymerization and condensation of receptors PNAS 2017 114 (50) 13188-13193; published ahead of print November 30, 2017, doi:10.1073/pnas.1713564114
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83607
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:30 Nov 2017 22:25
Last Modified:03 Oct 2019 19:08

Repository Staff Only: item control page