The sphingosine 1-phosphate receptor 2 is shed in exosomes from breast cancer cells and is N-terminally processed to a short constitutively active form that promotes extracellular signal regulated kinase activation and DNA synthesis in fibroblasts

Ashref El Buri, David R. Adams, Douglas Smith, Rothwelle J. Tate, Margaret Mullin, Susan Pyne, Nigel J. Pyne

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5 Citations (Scopus)

Abstract

We demonstrate here that the G protein-coupled receptor (GPCR), sphingosine 1-phosphate receptor 2 (S1P2, Mr = 40 kDa) is shed in hsp70+ and CD63+ containing exosomes from MDA-MB-231 breast cancer cells. The receptor is taken up by fibroblasts, where it is N-terminally processed to a shorter form (Mr = 36 kDa) that appears to be constitutively active and able to stimulate the extracellular signal regulated kinase-1/2 (ERK-1/2) pathway and DNA synthesis. An N-terminally truncated construct of S1P2, which may correspond to the processed form of the receptor generated in fibroblasts, was found to be constitutively active when over-expressed in HEK293 cells. Analysis based on the available crystal structure of the homologous S1P1 receptor suggests that, in the inactive-state, the N-terminus of S1P2 may tension TM1 so as to maintain a compressive action on TM7. This in turn may stabilise a closed basal state interface between the intracellular ends of TM7 and TM6. Cleavage and removal of the S1P2 N-terminal peptide is postulated to facilitate relaxation of TM1 and accompanying separation of TM6 and TM7. The latter transition is one of the key elements of G protein engagement and is required to open the intracellular coupling interface beneath the GPCR helix bundle. Therefore, removal at the N-terminus of S1P2 is likely to enhance G protein coupling. These findings provide the first evidence that S1P2 is released from breast cancer cells in exosomes and is processed by fibroblasts to promote ERK signaling and proliferation of these cells.
LanguageEnglish
Pages29453-29467
Number of pages15
JournalOncotarget
Volume9
Issue number50
DOIs
Publication statusPublished - 29 Jun 2018

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Lysosphingolipid Receptors
Exosomes
Extracellular Signal-Regulated MAP Kinases
Fibroblasts
Breast Neoplasms
G-Protein-Coupled Receptors
GTP-Binding Proteins
DNA
Mitogen-Activated Protein Kinase 3
HEK293 Cells
Mitogen-Activated Protein Kinase 1
Cell Proliferation
Peptides

Keywords

  • sphingosine 1-phosphate
  • sphingosine 1-phosphate receptor
  • exosomes
  • fibroblasts

Cite this

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title = "The sphingosine 1-phosphate receptor 2 is shed in exosomes from breast cancer cells and is N-terminally processed to a short constitutively active form that promotes extracellular signal regulated kinase activation and DNA synthesis in fibroblasts",
abstract = "We demonstrate here that the G protein-coupled receptor (GPCR), sphingosine 1-phosphate receptor 2 (S1P2, Mr = 40 kDa) is shed in hsp70+ and CD63+ containing exosomes from MDA-MB-231 breast cancer cells. The receptor is taken up by fibroblasts, where it is N-terminally processed to a shorter form (Mr = 36 kDa) that appears to be constitutively active and able to stimulate the extracellular signal regulated kinase-1/2 (ERK-1/2) pathway and DNA synthesis. An N-terminally truncated construct of S1P2, which may correspond to the processed form of the receptor generated in fibroblasts, was found to be constitutively active when over-expressed in HEK293 cells. Analysis based on the available crystal structure of the homologous S1P1 receptor suggests that, in the inactive-state, the N-terminus of S1P2 may tension TM1 so as to maintain a compressive action on TM7. This in turn may stabilise a closed basal state interface between the intracellular ends of TM7 and TM6. Cleavage and removal of the S1P2 N-terminal peptide is postulated to facilitate relaxation of TM1 and accompanying separation of TM6 and TM7. The latter transition is one of the key elements of G protein engagement and is required to open the intracellular coupling interface beneath the GPCR helix bundle. Therefore, removal at the N-terminus of S1P2 is likely to enhance G protein coupling. These findings provide the first evidence that S1P2 is released from breast cancer cells in exosomes and is processed by fibroblasts to promote ERK signaling and proliferation of these cells.",
keywords = "sphingosine 1-phosphate, sphingosine 1-phosphate receptor, exosomes, fibroblasts",
author = "{El Buri}, Ashref and Adams, {David R.} and Douglas Smith and Tate, {Rothwelle J.} and Margaret Mullin and Susan Pyne and Pyne, {Nigel J.}",
year = "2018",
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AU - El Buri, Ashref

AU - Adams, David R.

AU - Smith, Douglas

AU - Tate, Rothwelle J.

AU - Mullin, Margaret

AU - Pyne, Susan

AU - Pyne, Nigel J.

PY - 2018/6/29

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N2 - We demonstrate here that the G protein-coupled receptor (GPCR), sphingosine 1-phosphate receptor 2 (S1P2, Mr = 40 kDa) is shed in hsp70+ and CD63+ containing exosomes from MDA-MB-231 breast cancer cells. The receptor is taken up by fibroblasts, where it is N-terminally processed to a shorter form (Mr = 36 kDa) that appears to be constitutively active and able to stimulate the extracellular signal regulated kinase-1/2 (ERK-1/2) pathway and DNA synthesis. An N-terminally truncated construct of S1P2, which may correspond to the processed form of the receptor generated in fibroblasts, was found to be constitutively active when over-expressed in HEK293 cells. Analysis based on the available crystal structure of the homologous S1P1 receptor suggests that, in the inactive-state, the N-terminus of S1P2 may tension TM1 so as to maintain a compressive action on TM7. This in turn may stabilise a closed basal state interface between the intracellular ends of TM7 and TM6. Cleavage and removal of the S1P2 N-terminal peptide is postulated to facilitate relaxation of TM1 and accompanying separation of TM6 and TM7. The latter transition is one of the key elements of G protein engagement and is required to open the intracellular coupling interface beneath the GPCR helix bundle. Therefore, removal at the N-terminus of S1P2 is likely to enhance G protein coupling. These findings provide the first evidence that S1P2 is released from breast cancer cells in exosomes and is processed by fibroblasts to promote ERK signaling and proliferation of these cells.

AB - We demonstrate here that the G protein-coupled receptor (GPCR), sphingosine 1-phosphate receptor 2 (S1P2, Mr = 40 kDa) is shed in hsp70+ and CD63+ containing exosomes from MDA-MB-231 breast cancer cells. The receptor is taken up by fibroblasts, where it is N-terminally processed to a shorter form (Mr = 36 kDa) that appears to be constitutively active and able to stimulate the extracellular signal regulated kinase-1/2 (ERK-1/2) pathway and DNA synthesis. An N-terminally truncated construct of S1P2, which may correspond to the processed form of the receptor generated in fibroblasts, was found to be constitutively active when over-expressed in HEK293 cells. Analysis based on the available crystal structure of the homologous S1P1 receptor suggests that, in the inactive-state, the N-terminus of S1P2 may tension TM1 so as to maintain a compressive action on TM7. This in turn may stabilise a closed basal state interface between the intracellular ends of TM7 and TM6. Cleavage and removal of the S1P2 N-terminal peptide is postulated to facilitate relaxation of TM1 and accompanying separation of TM6 and TM7. The latter transition is one of the key elements of G protein engagement and is required to open the intracellular coupling interface beneath the GPCR helix bundle. Therefore, removal at the N-terminus of S1P2 is likely to enhance G protein coupling. These findings provide the first evidence that S1P2 is released from breast cancer cells in exosomes and is processed by fibroblasts to promote ERK signaling and proliferation of these cells.

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