NRas activity is regulated by dynamic interactions with nanoscale signaling clusters at the plasma membrane

Oren Yakovian; Julia Sajman; Michal Alon; Rand Arafeh; Yardena Samuels; Eilon Sherman

doi:10.1016/j.isci.2022.105282

NRas activity is regulated by dynamic interactions with nanoscale signaling clusters at the plasma membrane

Oren Yakovian, Julia Sajman, Michal Alon, Rand Arafeh, Yardena Samuels, Eilon Sherman^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

NRas is a key mediator of the mitogenic pathway in normal cells and in cancer cells. Its dynamics and nanoscale organization at the plasma membrane (PM) facilitate its signaling. Here, we used two-color photoactivated localization microscopy to resolve the organization of individual NRas and associated signaling proteins in live melanoma cells, with resolution down to ∼20 nm. Upon EGF activation, a fraction of NRas and BRAF (dis)assembled synchronously at the PM in co-clusters. NRas and BRAF clusters associated with GPI-enriched domains, serving as possible nucleation sites for these clusters. NRas and BRAF association in mutual clusters was reduced by the NRas farnesylation inhibitor lonafarnib, yet enhanced by the BRAF inhibitor vemurafenib. Surprisingly, dispersed NRas molecules associated with the periphery of self-clusters of either Grb2 or NF1. Thus, NRas-mediated signaling, which is critical in health and disease, is regulated by dynamic interactions with functional clusters of BRAF or other related proteins at the PM.

Original language	American English
Article number	105282
Journal	iScience
Volume	25
Issue number	11
DOIs	https://doi.org/10.1016/j.isci.2022.105282
State	Published - 18 Nov 2022

Bibliographical note

Funding Information:
This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17, the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 754282), the ERC (CoG- 770854), MRA (#622106), Rising Tide Foundation, Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics, Estate of Alice Schwarz-Gardos, Estate of John Hunter, Knell Family, Peter and Patricia Gruber Award, and the Hamburger Family. E.S. supervised the research. E.S. J.S. Y.S. R.A. and O.Y. designed the research; M.A. and J.S. assisted with critical reagents; O.Y. performed research; O.Y. analyzed the data; E.S. and O.Y. wrote the article; all co-authors commented on the article. The authors declare no potential conflicts of interest.

Funding Information:
This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17 , the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No. 754282 ), the ERC (CoG- 770854 ), MRA ( #622106 ), Rising Tide Foundation , Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics , Estate of Alice Schwarz-Gardos , Estate of John Hunter , Knell Family , Peter and Patricia Gruber Award, and the Hamburger Family .

Publisher Copyright:
© 2022

Keywords

Biological sciences
Biophysics
Cell biology
Molecular biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.isci.2022.105282

Cite this

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title = "NRas activity is regulated by dynamic interactions with nanoscale signaling clusters at the plasma membrane",

abstract = "NRas is a key mediator of the mitogenic pathway in normal cells and in cancer cells. Its dynamics and nanoscale organization at the plasma membrane (PM) facilitate its signaling. Here, we used two-color photoactivated localization microscopy to resolve the organization of individual NRas and associated signaling proteins in live melanoma cells, with resolution down to ∼20 nm. Upon EGF activation, a fraction of NRas and BRAF (dis)assembled synchronously at the PM in co-clusters. NRas and BRAF clusters associated with GPI-enriched domains, serving as possible nucleation sites for these clusters. NRas and BRAF association in mutual clusters was reduced by the NRas farnesylation inhibitor lonafarnib, yet enhanced by the BRAF inhibitor vemurafenib. Surprisingly, dispersed NRas molecules associated with the periphery of self-clusters of either Grb2 or NF1. Thus, NRas-mediated signaling, which is critical in health and disease, is regulated by dynamic interactions with functional clusters of BRAF or other related proteins at the PM.",

keywords = "Biological sciences, Biophysics, Cell biology, Molecular biology",

author = "Oren Yakovian and Julia Sajman and Michal Alon and Rand Arafeh and Yardena Samuels and Eilon Sherman",

note = "Funding Information: This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17, the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 754282), the ERC (CoG- 770854), MRA (#622106), Rising Tide Foundation, Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics, Estate of Alice Schwarz-Gardos, Estate of John Hunter, Knell Family, Peter and Patricia Gruber Award, and the Hamburger Family. E.S. supervised the research. E.S. J.S. Y.S. R.A. and O.Y. designed the research; M.A. and J.S. assisted with critical reagents; O.Y. performed research; O.Y. analyzed the data; E.S. and O.Y. wrote the article; all co-authors commented on the article. The authors declare no potential conflicts of interest. Funding Information: This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17 , the European Research Council (ERC) under the European Union{\textquoteright}s Horizon 2020 research and innovation programme (grant agreement No. 754282 ), the ERC (CoG- 770854 ), MRA ( #622106 ), Rising Tide Foundation , Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics , Estate of Alice Schwarz-Gardos , Estate of John Hunter , Knell Family , Peter and Patricia Gruber Award, and the Hamburger Family . Publisher Copyright: {\textcopyright} 2022",

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AU - Sajman, Julia

AU - Alon, Michal

AU - Arafeh, Rand

AU - Samuels, Yardena

AU - Sherman, Eilon

N1 - Funding Information: This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17, the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 754282), the ERC (CoG- 770854), MRA (#622106), Rising Tide Foundation, Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics, Estate of Alice Schwarz-Gardos, Estate of John Hunter, Knell Family, Peter and Patricia Gruber Award, and the Hamburger Family. E.S. supervised the research. E.S. J.S. Y.S. R.A. and O.Y. designed the research; M.A. and J.S. assisted with critical reagents; O.Y. performed research; O.Y. analyzed the data; E.S. and O.Y. wrote the article; all co-authors commented on the article. The authors declare no potential conflicts of interest. Funding Information: This research was supported by Grants no. 1761/17 and no. 1937/13 from the Israel Science Foundation (E.S.). Y.S. is supported by the Israel Science Foundation grant No. 696/17 , the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No. 754282 ), the ERC (CoG- 770854 ), MRA ( #622106 ), Rising Tide Foundation , Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics , Estate of Alice Schwarz-Gardos , Estate of John Hunter , Knell Family , Peter and Patricia Gruber Award, and the Hamburger Family . Publisher Copyright: © 2022

PY - 2022/11/18

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N2 - NRas is a key mediator of the mitogenic pathway in normal cells and in cancer cells. Its dynamics and nanoscale organization at the plasma membrane (PM) facilitate its signaling. Here, we used two-color photoactivated localization microscopy to resolve the organization of individual NRas and associated signaling proteins in live melanoma cells, with resolution down to ∼20 nm. Upon EGF activation, a fraction of NRas and BRAF (dis)assembled synchronously at the PM in co-clusters. NRas and BRAF clusters associated with GPI-enriched domains, serving as possible nucleation sites for these clusters. NRas and BRAF association in mutual clusters was reduced by the NRas farnesylation inhibitor lonafarnib, yet enhanced by the BRAF inhibitor vemurafenib. Surprisingly, dispersed NRas molecules associated with the periphery of self-clusters of either Grb2 or NF1. Thus, NRas-mediated signaling, which is critical in health and disease, is regulated by dynamic interactions with functional clusters of BRAF or other related proteins at the PM.

AB - NRas is a key mediator of the mitogenic pathway in normal cells and in cancer cells. Its dynamics and nanoscale organization at the plasma membrane (PM) facilitate its signaling. Here, we used two-color photoactivated localization microscopy to resolve the organization of individual NRas and associated signaling proteins in live melanoma cells, with resolution down to ∼20 nm. Upon EGF activation, a fraction of NRas and BRAF (dis)assembled synchronously at the PM in co-clusters. NRas and BRAF clusters associated with GPI-enriched domains, serving as possible nucleation sites for these clusters. NRas and BRAF association in mutual clusters was reduced by the NRas farnesylation inhibitor lonafarnib, yet enhanced by the BRAF inhibitor vemurafenib. Surprisingly, dispersed NRas molecules associated with the periphery of self-clusters of either Grb2 or NF1. Thus, NRas-mediated signaling, which is critical in health and disease, is regulated by dynamic interactions with functional clusters of BRAF or other related proteins at the PM.

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SN - 2589-0042

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NRas activity is regulated by dynamic interactions with nanoscale signaling clusters at the plasma membrane

Abstract

Bibliographical note

Keywords

UN SDGs

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