Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force

Ioanna Smyrlaki; Ferenc Fördős; Iris Rocamonde-Lago; Yang Wang; Boxuan Shen; Antonio Lentini; Vincent C. Luca; Björn Reinius; Ana I. Teixeira; Björn Högberg

doi:10.1038/s41467-023-44059-4

Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force

Ioanna Smyrlaki
, Ferenc Fördős
, Iris Rocamonde-Lago
, Yang Wang
, Boxuan Shen
, Antonio Lentini
, Vincent C. Luca
, Björn Reinius
, Ana I. Teixeira
, Björn Högberg^*

^*Corresponding author for this work

Karolinska Institutet
Moffitt Cancer Center

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

25 Citations (Scopus)

44 Downloads (Pure)

Abstract

The Notch signaling pathway has fundamental roles in embryonic development and in the nervous system. The current model of receptor activation involves initiation via a force-induced conformational change. Here, we define conditions that reveal pulling force-independent Notch activation using soluble multivalent constructs. We treat neuroepithelial stem-like cells with molecularly precise ligand nanopatterns displayed from solution using DNA origami. Notch signaling follows with clusters of Jag1, and with chimeric structures where most Jag1 proteins are replaced by other binders not targeting Notch. Our data rule out several confounding factors and suggest a model where Jag1 activates Notch upon prolonged binding without appearing to need a pulling force. These findings reveal a distinct mode of activation of Notch and lay the foundation for the development of soluble agonists.

Original language	English
Article number	465
Number of pages	14
Journal	Nature Communications
Volume	15
Issue number	1
DOIs	https://doi.org/10.1038/s41467-023-44059-4
Publication status	Published - 18 Jan 2024
MoE publication type	A1 Journal article-refereed

Funding

The authors would like to acknowledge support from the NIH grant number R35GM133482 for V.C.L., the Knut and Alice Wallenberg Foundation (Grants KAW 2017.0114 for B.H. and A.I.T. and KAW 2017.0276 for B.H.), from the European Research Council ERC for B.H. (Acronym: Cell Track GA No. 724872) and A.I.T (Acronym: MechComm GA No. 617711), and from the Swedish Research Council for B.H. (grant no. 2019-01474) and from the Göran Gustafsson Foundation for B.H. And from the Academy of Finland for B.S. (grant no. 341908). lt-NES samples were obtained from, and initial culture protocols was made possible with the help of Anna Falk’s team and the iPS Core facility at Karolinska Institutet. Part of this work was performed at the Karolinska Institutet/SciLifeLab Protein Science Core Facility (PSF). Part of this work was performed at the Karolinska Institutet Biomedicum Imaging Core (BIC). EM data was collected at the Karolinska Institutet 3D-EM facility. The authors would like to acknowledge support from the NIH grant number R35GM133482 for V.C.L., the Knut and Alice Wallenberg Foundation (Grants KAW 2017.0114 for B.H. and A.I.T. and KAW 2017.0276 for B.H.), from the European Research Council ERC for B.H. (Acronym: Cell Track GA No. 724872) and A.I.T (Acronym: MechComm GA No. 617711), and from the Swedish Research Council for B.H. (grant no. 2019-01474) and from the Göran Gustafsson Foundation for B.H. And from the Academy of Finland for B.S. (grant no. 341908). lt-NES samples were obtained from, and initial culture protocols was made possible with the help of Anna Falk’s team and the iPS Core facility at Karolinska Institutet. Part of this work was performed at the Karolinska Institutet/SciLifeLab Protein Science Core Facility (PSF). Part of this work was performed at the Karolinska Institutet Biomedicum Imaging Core (BIC). EM data was collected at the Karolinska Institutet 3D-EM facility.

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Cite this

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title = "Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force",

abstract = "The Notch signaling pathway has fundamental roles in embryonic development and in the nervous system. The current model of receptor activation involves initiation via a force-induced conformational change. Here, we define conditions that reveal pulling force-independent Notch activation using soluble multivalent constructs. We treat neuroepithelial stem-like cells with molecularly precise ligand nanopatterns displayed from solution using DNA origami. Notch signaling follows with clusters of Jag1, and with chimeric structures where most Jag1 proteins are replaced by other binders not targeting Notch. Our data rule out several confounding factors and suggest a model where Jag1 activates Notch upon prolonged binding without appearing to need a pulling force. These findings reveal a distinct mode of activation of Notch and lay the foundation for the development of soluble agonists.",

author = "Ioanna Smyrlaki and Ferenc F{\"o}rd{\H o}s and Iris Rocamonde-Lago and Yang Wang and Boxuan Shen and Antonio Lentini and Luca, \{Vincent C.\} and Bj{\"o}rn Reinius and Teixeira, \{Ana I.\} and Bj{\"o}rn H{\"o}gberg",

note = "Funding Information: The authors would like to acknowledge support from the NIH grant number R35GM133482 for V.C.L., the Knut and Alice Wallenberg Foundation (Grants KAW 2017.0114 for B.H. and A.I.T. and KAW 2017.0276 for B.H.), from the European Research Council ERC for B.H. (Acronym: Cell Track GA No. 724872) and A.I.T (Acronym: MechComm GA No. 617711), and from the Swedish Research Council for B.H. (grant no. 2019-01474) and from the G{\"o}ran Gustafsson Foundation for B.H. And from the Academy of Finland for B.S. (grant no. 341908). lt-NES samples were obtained from, and initial culture protocols was made possible with the help of Anna Falk{\textquoteright}s team and the iPS Core facility at Karolinska Institutet. Part of this work was performed at the Karolinska Institutet/SciLifeLab Protein Science Core Facility (PSF). Part of this work was performed at the Karolinska Institutet Biomedicum Imaging Core (BIC). EM data was collected at the Karolinska Institutet 3D-EM facility. Publisher Copyright: {\textcopyright} 2024, The Author(s).",

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AU - Smyrlaki, Ioanna

AU - Fördős, Ferenc

AU - Rocamonde-Lago, Iris

AU - Wang, Yang

AU - Shen, Boxuan

AU - Lentini, Antonio

AU - Luca, Vincent C.

AU - Reinius, Björn

AU - Teixeira, Ana I.

AU - Högberg, Björn

N1 - Funding Information: The authors would like to acknowledge support from the NIH grant number R35GM133482 for V.C.L., the Knut and Alice Wallenberg Foundation (Grants KAW 2017.0114 for B.H. and A.I.T. and KAW 2017.0276 for B.H.), from the European Research Council ERC for B.H. (Acronym: Cell Track GA No. 724872) and A.I.T (Acronym: MechComm GA No. 617711), and from the Swedish Research Council for B.H. (grant no. 2019-01474) and from the Göran Gustafsson Foundation for B.H. And from the Academy of Finland for B.S. (grant no. 341908). lt-NES samples were obtained from, and initial culture protocols was made possible with the help of Anna Falk’s team and the iPS Core facility at Karolinska Institutet. Part of this work was performed at the Karolinska Institutet/SciLifeLab Protein Science Core Facility (PSF). Part of this work was performed at the Karolinska Institutet Biomedicum Imaging Core (BIC). EM data was collected at the Karolinska Institutet 3D-EM facility. Publisher Copyright: © 2024, The Author(s).

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Y1 - 2024/1/18

N2 - The Notch signaling pathway has fundamental roles in embryonic development and in the nervous system. The current model of receptor activation involves initiation via a force-induced conformational change. Here, we define conditions that reveal pulling force-independent Notch activation using soluble multivalent constructs. We treat neuroepithelial stem-like cells with molecularly precise ligand nanopatterns displayed from solution using DNA origami. Notch signaling follows with clusters of Jag1, and with chimeric structures where most Jag1 proteins are replaced by other binders not targeting Notch. Our data rule out several confounding factors and suggest a model where Jag1 activates Notch upon prolonged binding without appearing to need a pulling force. These findings reveal a distinct mode of activation of Notch and lay the foundation for the development of soluble agonists.

AB - The Notch signaling pathway has fundamental roles in embryonic development and in the nervous system. The current model of receptor activation involves initiation via a force-induced conformational change. Here, we define conditions that reveal pulling force-independent Notch activation using soluble multivalent constructs. We treat neuroepithelial stem-like cells with molecularly precise ligand nanopatterns displayed from solution using DNA origami. Notch signaling follows with clusters of Jag1, and with chimeric structures where most Jag1 proteins are replaced by other binders not targeting Notch. Our data rule out several confounding factors and suggest a model where Jag1 activates Notch upon prolonged binding without appearing to need a pulling force. These findings reveal a distinct mode of activation of Notch and lay the foundation for the development of soluble agonists.

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DO - 10.1038/s41467-023-44059-4

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JO - Nature Communications

JF - Nature Communications

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M1 - 465

ER -

Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force

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SiDNAno/Shen: Silica-DNA Hybrid Nanostructures as Next Generation Self-Assembling Nanomaterials

Unfolding Mysteries: DNA Origami Unveils Hidden Mechanisms of Critical Cell Function

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Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force

Abstract

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SiDNAno/Shen: Silica-DNA Hybrid Nanostructures as Next Generation Self-Assembling Nanomaterials

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Unfolding Mysteries: DNA Origami Unveils Hidden Mechanisms of Critical Cell Function

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