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Distinct mechanisms of microRNA sorting into cancer cell-derived extracellular vesicle subtypes

View ORCID ProfileMorayma M. Temoche-Diaz, View ORCID ProfileMatthew J. Shurtleff, View ORCID ProfileRyan M. Nottingham, Jun Yao, Raj P. Fadadu, Alan M. Lambowitz, View ORCID ProfileRandy Schekman
doi: https://doi.org/10.1101/612069
Morayma M. Temoche-Diaz
1Department of Plant and Microbial Biology, University of California, Berkeley, United States
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Matthew J. Shurtleff
1Department of Plant and Microbial Biology, University of California, Berkeley, United States
2Department of Molecular and Cellular Biology, Howard Hughes Medical Institute, University of California, Berkeley, United States
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Ryan M. Nottingham
3Institute for Cellular and Molecular Biology and Department of Molecular Biosciences, University of Texas, Austin, United States
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Jun Yao
3Institute for Cellular and Molecular Biology and Department of Molecular Biosciences, University of Texas, Austin, United States
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Raj P. Fadadu
2Department of Molecular and Cellular Biology, Howard Hughes Medical Institute, University of California, Berkeley, United States
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Alan M. Lambowitz
3Institute for Cellular and Molecular Biology and Department of Molecular Biosciences, University of Texas, Austin, United States
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Randy Schekman
2Department of Molecular and Cellular Biology, Howard Hughes Medical Institute, University of California, Berkeley, United States
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  • For correspondence: schekman@berkeley.edu
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ABSTRACT

Extracellular vesicles (EVs) encompass a variety of vesicles secreted into the extracellular space. EVs have been implicated in promoting tumor metastasis, but the molecular composition of tumor-derived EV sub-types and the mechanisms by which molecules are sorted into EVs remain mostly unknown. We report the separation of two EV sub-populations from a metastatic breast cancer cell line, with biochemical features consistent with different sub-cellular origins. These EV sub-types use different mechanisms of miRNA sorting (selective and non-selective), suggesting that sorting occurs via fundamentally distinct processes, possibly dependent on EV origin. Using biochemical and genetic tools, we identified the Lupus La protein as mediating sorting of some selectively packaged miRNAs. We found that two motifs embedded in miR-122 are responsible for high-affinity binding to Lupus La and sorting into vesicles formed in a cell-free reaction. Thus, tumor cells can simultaneously deploy multiple EV species using distinct sorting mechanisms that may enable diverse functions in normal and cancer biology.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted April 17, 2019.
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Distinct mechanisms of microRNA sorting into cancer cell-derived extracellular vesicle subtypes
Morayma M. Temoche-Diaz, Matthew J. Shurtleff, Ryan M. Nottingham, Jun Yao, Raj P. Fadadu, Alan M. Lambowitz, Randy Schekman
bioRxiv 612069; doi: https://doi.org/10.1101/612069
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Distinct mechanisms of microRNA sorting into cancer cell-derived extracellular vesicle subtypes
Morayma M. Temoche-Diaz, Matthew J. Shurtleff, Ryan M. Nottingham, Jun Yao, Raj P. Fadadu, Alan M. Lambowitz, Randy Schekman
bioRxiv 612069; doi: https://doi.org/10.1101/612069

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