Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation

Abstract B‐cell receptor (BCR) signaling is essential for the development and function of B cells; however, the spectrum of proteins involved in BCR signaling is not fully known. Here we used quantitative mass spectrometry‐based proteomics to monitor the dynamics of BCR signaling complexes (signalos...

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Main Authors: Shankha Satpathy, Sebastian A Wagner, Petra Beli, Rajat Gupta, Trine A Kristiansen, Dessislava Malinova, Chiara Francavilla, Pavel Tolar, Gail A Bishop, Bruce S Hostager, Chunaram Choudhary
Format: Article
Language:English
Published: Springer Nature 2015-06-01
Series:Molecular Systems Biology
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Online Access:https://doi.org/10.15252/msb.20145880
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author Shankha Satpathy
Sebastian A Wagner
Petra Beli
Rajat Gupta
Trine A Kristiansen
Dessislava Malinova
Chiara Francavilla
Pavel Tolar
Gail A Bishop
Bruce S Hostager
Chunaram Choudhary
author_facet Shankha Satpathy
Sebastian A Wagner
Petra Beli
Rajat Gupta
Trine A Kristiansen
Dessislava Malinova
Chiara Francavilla
Pavel Tolar
Gail A Bishop
Bruce S Hostager
Chunaram Choudhary
author_sort Shankha Satpathy
collection DOAJ
description Abstract B‐cell receptor (BCR) signaling is essential for the development and function of B cells; however, the spectrum of proteins involved in BCR signaling is not fully known. Here we used quantitative mass spectrometry‐based proteomics to monitor the dynamics of BCR signaling complexes (signalosomes) and to investigate the dynamics of downstream phosphorylation and ubiquitylation signaling. We identify most of the previously known components of BCR signaling, as well as many proteins that have not yet been implicated in this system. BCR activation leads to rapid tyrosine phosphorylation and ubiquitylation of the receptor‐proximal signaling components, many of which are co‐regulated by both the modifications. We illustrate the power of multilayered proteomic analyses for discovering novel BCR signaling components by demonstrating that BCR‐induced phosphorylation of RAB7A at S72 prevents its association with effector proteins and with endo‐lysosomal compartments. In addition, we show that BCL10 is modified by LUBAC‐mediated linear ubiquitylation, and demonstrate an important function of LUBAC in BCR‐induced NF‐κB signaling. Our results offer a global and integrated view of BCR signaling, and the provided datasets can serve as a valuable resource for further understanding BCR signaling networks.
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spelling doaj-art-6e9e558b60ec42e8b4adc1da130fe1df2025-08-20T03:46:32ZengSpringer NatureMolecular Systems Biology1744-42922015-06-0111612210.15252/msb.20145880Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylationShankha Satpathy0Sebastian A Wagner1Petra Beli2Rajat Gupta3Trine A Kristiansen4Dessislava Malinova5Chiara Francavilla6Pavel Tolar7Gail A Bishop8Bruce S Hostager9Chunaram Choudhary10Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDivision of Immune Cell Biology, MRC National Institute for Medical ResearchDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenDivision of Immune Cell Biology, MRC National Institute for Medical ResearchDepartment of Microbiology, Graduate Program in Immunology and Department of Internal Medicine, University of IowaDepartment of Pediatrics, University of IowaDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenAbstract B‐cell receptor (BCR) signaling is essential for the development and function of B cells; however, the spectrum of proteins involved in BCR signaling is not fully known. Here we used quantitative mass spectrometry‐based proteomics to monitor the dynamics of BCR signaling complexes (signalosomes) and to investigate the dynamics of downstream phosphorylation and ubiquitylation signaling. We identify most of the previously known components of BCR signaling, as well as many proteins that have not yet been implicated in this system. BCR activation leads to rapid tyrosine phosphorylation and ubiquitylation of the receptor‐proximal signaling components, many of which are co‐regulated by both the modifications. We illustrate the power of multilayered proteomic analyses for discovering novel BCR signaling components by demonstrating that BCR‐induced phosphorylation of RAB7A at S72 prevents its association with effector proteins and with endo‐lysosomal compartments. In addition, we show that BCL10 is modified by LUBAC‐mediated linear ubiquitylation, and demonstrate an important function of LUBAC in BCR‐induced NF‐κB signaling. Our results offer a global and integrated view of BCR signaling, and the provided datasets can serve as a valuable resource for further understanding BCR signaling networks.https://doi.org/10.15252/msb.20145880BCL10BCRphosphorylationRAB7Aubiquitylation
spellingShingle Shankha Satpathy
Sebastian A Wagner
Petra Beli
Rajat Gupta
Trine A Kristiansen
Dessislava Malinova
Chiara Francavilla
Pavel Tolar
Gail A Bishop
Bruce S Hostager
Chunaram Choudhary
Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
Molecular Systems Biology
BCL10
BCR
phosphorylation
RAB7A
ubiquitylation
title Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
title_full Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
title_fullStr Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
title_full_unstemmed Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
title_short Systems‐wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation
title_sort systems wide analysis of bcr signalosomes and downstream phosphorylation and ubiquitylation
topic BCL10
BCR
phosphorylation
RAB7A
ubiquitylation
url https://doi.org/10.15252/msb.20145880
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