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  Condensed-phase signaling can expand kinase specificity and respond to macromolecular crowding

Sang, D., Shu, T., Pantoja, C. F., de Opakua, A. I., Zweckstetter, M., & Holt, L. J. (2022). Condensed-phase signaling can expand kinase specificity and respond to macromolecular crowding. Molecular Cell, 82(19), 3693-3711.e10. doi:10.1016/j.molcel.2022.08.016.

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 Creators:
Sang, Dajun, Author
Shu, Tong, Author
Pantoja, Christian F., Author
de Opakua, Alain Ibáñez, Author
Zweckstetter, M.1, Author           
Holt, Liam J., Author
Affiliations:
1Department of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, Göttingen, DE, ou_3350124              

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 Abstract: Phase separation can concentrate biomolecules and accelerate reactions. However, the mechanisms and principles connecting this mesoscale organization to signaling dynamics are difficult to dissect because of the pleiotropic effects associated with disrupting endogenous condensates. To address this limitation, we engineered new phosphorylation reactions within synthetic condensates. We generally found increased activity and broadened kinase specificity. Phosphorylation dynamics within condensates were rapid and could drive cell-cycle-dependent localization changes. High client concentration within condensates was important but not the main factor for efficient phosphorylation. Rather, the availability of many excess client-binding sites together with a flexible scaffold was crucial. Phosphorylation within condensates was also modulated by changes in macromolecular crowding. Finally, the phosphorylation of the Alzheimer’s-disease-associated protein Tau by cyclin-dependent kinase 2 was accelerated within condensates. Thus, condensates enable new signaling connections and can create sensors that respond to the biophysical properties of the cytoplasm.

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Language(s): eng - English
 Dates: 2022-09-142022-10-06
 Publication Status: Issued
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.molcel.2022.08.016
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Project name : We thank Michael Rosen, Allyson Rice, and members of the HHMI Summer Institute at the Marine Biology Institute in Woods Hole for discussions that initiated this project. We thank Jef Boeke, David Engelberg, Meta Heidenreich, and Emmanuel D. Levy for sharing yeast strains and plasmids, and Greg Brittingham for help with imaging and analysis. We thank Lance Denes and Srinjoy Sil for critical reading of the manuscript. We thank the rest of the Holt lab for helpful discussions. We thank Maria-Sol Cima-Omori for preparation of 15N-labeled Tau. C.F.P. acknowledges the Deutsche Akademischer Austauschdienst (DAAD) for the Research Grants Doctoral program in Germany, 2019/20 (personal ref. no. 91726791) and M.Z. was supported by the European Research Council (ERC) under the EU Horizon 2020 research and innovation program (grant agreement no. 787679). L.J.H. was funded by NIH R01 GM132447 and R37 CA240765, a donation to the American Cancer Society from the Cornelia T. Bailey Foundation, the Pershing Square Sohn Cancer Research Award, the Chan Zuckerberg Initiative, the NIH Director’s Transformative Research Award TR01 NS127186, and the Air Force Office of Scientific Research (AFoSR) Multidisciplinary University Research Initiative (MURI) competition (grant FA9550-21-1-3503 0091).
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Grant ID : 787679
Funding program : Horizon 2020 (H2020)
Funding organization : European Commission (EC)

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Title: Molecular Cell
Source Genre: Journal
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Publ. Info: Cambridge, Mass. : Cell Press
Pages: - Volume / Issue: 82 (19) Sequence Number: - Start / End Page: 3693 - 3711.e10 Identifier: ISSN: 1097-2765
CoNE: https://pure.mpg.de/cone/journals/resource/954925610929