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

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A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

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en

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14

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Nature Communications, Volume 15, issue 1

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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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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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Smyrlaki, I, Fördős, F, Rocamonde-Lago, I, Wang, Y, Shen, B, Lentini, A, Luca, V C, Reinius, B, Teixeira, A I & Högberg, B 2024, 'Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force', Nature Communications, vol. 15, no. 1, 465. https://doi.org/10.1038/s41467-023-44059-4