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Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling

Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite that serves as a potent extracellular signaling molecule. Metabolic regulation of extracellular S1P levels impacts key cellular activities through altered S1P receptor signaling. Although the pathway through which S1P is degraded within the...

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Autores principales: Kono, Mari, Hoachlander-Hobby, Lila E., Majumder, Saurav, Schwartz, Ronit, Byrnes, Colleen, Zhu, Hongling, Proia, Richard L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9213771/
https://www.ncbi.nlm.nih.gov/pubmed/35568252
http://dx.doi.org/10.1016/j.jlr.2022.100225
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author Kono, Mari
Hoachlander-Hobby, Lila E.
Majumder, Saurav
Schwartz, Ronit
Byrnes, Colleen
Zhu, Hongling
Proia, Richard L.
author_facet Kono, Mari
Hoachlander-Hobby, Lila E.
Majumder, Saurav
Schwartz, Ronit
Byrnes, Colleen
Zhu, Hongling
Proia, Richard L.
author_sort Kono, Mari
collection PubMed
description Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite that serves as a potent extracellular signaling molecule. Metabolic regulation of extracellular S1P levels impacts key cellular activities through altered S1P receptor signaling. Although the pathway through which S1P is degraded within the cell and thereby eliminated from reuse has been previously described, the mechanism used for S1P cellular uptake and the subsequent recycling of its sphingoid base into the sphingolipid synthesis pathway is not completely understood. To identify the genes within this S1P uptake and recycling pathway, we performed a genome-wide CRISPR/Cas9 KO screen using a positive-selection scheme with Shiga toxin, which binds a cell-surface glycosphingolipid receptor, globotriaosylceramide (Gb3), and causes lethality upon internalization. The screen was performed in HeLa cells with their sphingolipid de novo pathway disabled so that Gb3 cell-surface expression was dependent on salvage of the sphingoid base of S1P taken up from the medium. The screen identified a suite of genes necessary for S1P uptake and the recycling of its sphingoid base to synthesize Gb3, including two lipid phosphatases, PLPP3 (phospholipid phosphatase 3) and SGPP1 (S1P phosphatase 1). The results delineate a pathway in which plasma membrane–bound PLPP3 dephosphorylates extracellular S1P to sphingosine, which then enters cells and is rephosphorylated to S1P by the sphingosine kinases. This rephosphorylation step is important to regenerate intracellular S1P as a branch-point substrate that can be routed either for dephosphorylation to salvage sphingosine for recycling into complex sphingolipid synthesis or for degradation to remove it from the sphingolipid synthesis pathway.
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spelling pubmed-92137712022-06-29 Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling Kono, Mari Hoachlander-Hobby, Lila E. Majumder, Saurav Schwartz, Ronit Byrnes, Colleen Zhu, Hongling Proia, Richard L. J Lipid Res Research Article Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite that serves as a potent extracellular signaling molecule. Metabolic regulation of extracellular S1P levels impacts key cellular activities through altered S1P receptor signaling. Although the pathway through which S1P is degraded within the cell and thereby eliminated from reuse has been previously described, the mechanism used for S1P cellular uptake and the subsequent recycling of its sphingoid base into the sphingolipid synthesis pathway is not completely understood. To identify the genes within this S1P uptake and recycling pathway, we performed a genome-wide CRISPR/Cas9 KO screen using a positive-selection scheme with Shiga toxin, which binds a cell-surface glycosphingolipid receptor, globotriaosylceramide (Gb3), and causes lethality upon internalization. The screen was performed in HeLa cells with their sphingolipid de novo pathway disabled so that Gb3 cell-surface expression was dependent on salvage of the sphingoid base of S1P taken up from the medium. The screen identified a suite of genes necessary for S1P uptake and the recycling of its sphingoid base to synthesize Gb3, including two lipid phosphatases, PLPP3 (phospholipid phosphatase 3) and SGPP1 (S1P phosphatase 1). The results delineate a pathway in which plasma membrane–bound PLPP3 dephosphorylates extracellular S1P to sphingosine, which then enters cells and is rephosphorylated to S1P by the sphingosine kinases. This rephosphorylation step is important to regenerate intracellular S1P as a branch-point substrate that can be routed either for dephosphorylation to salvage sphingosine for recycling into complex sphingolipid synthesis or for degradation to remove it from the sphingolipid synthesis pathway. American Society for Biochemistry and Molecular Biology 2022-05-11 /pmc/articles/PMC9213771/ /pubmed/35568252 http://dx.doi.org/10.1016/j.jlr.2022.100225 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Kono, Mari
Hoachlander-Hobby, Lila E.
Majumder, Saurav
Schwartz, Ronit
Byrnes, Colleen
Zhu, Hongling
Proia, Richard L.
Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title_full Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title_fullStr Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title_full_unstemmed Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title_short Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
title_sort identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9213771/
https://www.ncbi.nlm.nih.gov/pubmed/35568252
http://dx.doi.org/10.1016/j.jlr.2022.100225
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