Abstract
Tonic or chronic active B-cell receptor (BCR) signaling is essential for the survival of normal or some malignant B cells, respectively. However, the molecular mechanism regulating the strength of these two types of BCR signaling remains unknown. Here, using high-speed high-resolution single-molecule tracking in live cells, we identified that PKCĪ², STIM1, and IP3R1/2/3 molecules affected the lateral Brownian mobile behavior of BCRs on the plasma membrane of quiescent B cells, which was correlated to the strength of BCR signaling. Further mechanistic studies revealed that these three molecules influenced BCR mobility by regulating the membrane tethering of MARCKS to the inner leaflet of the plasma membrane. Indeed, membrane-untethered or deficiency of MARCKS significantly decreased, while membrane-tethered or overexpression of MARCKS drastically increased the lateral mobility of BCRs. Functional experiments indicated that the membrane-tethered MARCKS suppressed the survival and/or proliferation in both B-cell tumor cells and mouse primary splenic B cells in vitro and in vivo. Mechanistically, we found that membrane-tethered MARCKS increased BCR lateral mobility, and thus decreased BCR nanoclustering by disturbing the interaction between cortical F-actin and the inner leaflet of the plasma membrane, resulting in the suppression of the strength of both tonic and chronic active BCR signaling. Conclusively, MARCKS is a newly identified molecule regulating the strength of BCR signaling by modulating cytoskeleton and plasma membrane interactions, both in the physiological and pathological conditions, suggesting thatĀ MARCKS is a putative target for drug design.
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Change history
01 December 2023
A Correction to this paper has been published: https://doi.org/10.1038/s41375-023-02091-9
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Acknowledgements
We thank Dr. Tomohiro Kurosaki and Dr. Hisaaki Shinohara (WPI Immunology Frontier Research Center, Osaka University, Japan) for providing DT40 cell lines, Dr. Carlos G. Dotti (Consejo Superior de Investigaciones CientĆficas) for providing MARCKS plasmid, Dr. Feng Zhang (MIT, Cambridge) for providing plasmids of pSpCas9-2a-GFP. We are supported by funds from National Science Foundation China (8182500030, 81730043 and 81621002) and Ministry of Science and Technology of China (2014CB542500-03).
Author Contributions
CX, YF and YJ performed the experiments and statistical analysis. CX and WL conceived the project. WL supervised the project. YZ, CL and YZ provided suggestions. CX and WL wrote the manuscript.
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Xu, C., Fang, Y., Yang, Z. et al. MARCKS regulates tonic and chronic active B cell receptor signaling. Leukemia 33, 710ā729 (2019). https://doi.org/10.1038/s41375-018-0244-4
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DOI: https://doi.org/10.1038/s41375-018-0244-4
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