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@article{1357368,
author = {Haugsten, Ellen Margrethe and Sørensen, Vigdis and Bosáková, Michaela and Souza, Gustavo Antonio de and Krejčí, Pavel and Wiedlocha, Antoni and Wesche, Jørgen},
article_location = {Washington},
article_number = {10},
doi = {http://dx.doi.org/10.1021/acs.jproteome.6b00652},
keywords = {FGFR4; FGF1; BioID; quantitative MS; confocal microscopy; three-dimensional structured illumination microscopy; endocytosis; clathrin; recycling compartment; trans-Golgi network},
language = {eng},
issn = {1535-3893},
journal = {Journal of Proteome Research},
title = {Proximity Labeling Reveals Molecular Determinants of FGFR4 Endosomal Transport},
volume = {15},
year = {2016}
}
TY - JOUR
ID - 1357368
AU - Haugsten, Ellen Margrethe - Sørensen, Vigdis - Bosáková, Michaela - Souza, Gustavo Antonio de - Krejčí, Pavel - Wiedlocha, Antoni - Wesche, Jørgen
PY - 2016
TI - Proximity Labeling Reveals Molecular Determinants of FGFR4 Endosomal Transport
JF - Journal of Proteome Research
VL - 15
IS - 10
SP - 3841-3855
EP - 3841-3855
PB - American Chemical Society
SN - 15353893
KW - FGFR4
KW - FGF1
KW - BioID
KW - quantitative MS
KW - confocal microscopy
KW - three-dimensional structured illumination microscopy
KW - endocytosis
KW - clathrin
KW - recycling compartment
KW - trans-Golgi network
N2 - The fibroblast growth factor receptors (FGFRs) are important oncogenes promoting tumor progression in many types of cancer, such as breast, bladder, and lung cancer as well as multiple myeloma and rhabdomyosarcoma. However, little is known about how these receptors are internalized and down-regulated in cells. We have here applied proximity biotin labeling to identify proteins involved in FGFR4 signaling and trafficking. For this purpose we fused a mutated biotin ligase, BirA*, to the C-terminal tail of FGFR4 (FGFR4-BirA*) and the fusion protein was stably expressed in U2OS cells. Upon addition of biotin to these cells, proteins in proximity to the FGFR4-BirA* fusion protein became biotinylated and could be isolated and identified by quantitative mass spectrometry. We identified in total 291 proteins, including 80 proteins that were enriched in samples where the receptor was activated by the ligand (FGF1), among them several proteins previously found to be involved in FGFR signaling (e.g., FRS2, PLC gamma, RSK2 and NCK2). Interestingly, many of the identified proteins were implicated in endosomal transport, and by precise annotation we were able to trace the intracellular pathways of activated FGFR4. Validating the data by confocal and three-dimensional structured illumination microscopy analysis, we concluded that FGFR4 uses clathrin-mediated endocytosis for internalization and is further sorted from early endosomes to the recycling compartment and the trans-Golgi network. Depletion of cells for clathrin heavy chain led to accumulation of FGFR4 at the cell surface and increased levels of active FGFR4 and PLC?, while AKT and ERK signaling was diminished, demonstrating that functional clathrin-mediated endocytosis is required for proper FGFR4 signaling. Thus, this study reveals proteins and pathways involved in FGFR4 transport and signaling that provide possible targets and opportunities for therapeutic intervention in FGFR4 aberrant cancer.
ER -
HAUGSTEN, Ellen Margrethe, Vigdis SØRENSEN, Michaela BOSÁKOVÁ, Gustavo Antonio de SOUZA, Pavel KREJČÍ, Antoni WIEDLOCHA and Jørgen WESCHE. Proximity Labeling Reveals Molecular Determinants of FGFR4 Endosomal Transport. \textit{Journal of Proteome Research}. Washington: American Chemical Society, 2016, vol.~15, No~10, p.~3841-3855. ISSN~1535-3893. Available from: https://dx.doi.org/10.1021/acs.jproteome.6b00652.
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