title: 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 (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.
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Shankha
authors.initials: S
authors.lastname: Satpathy
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Sebastian A
authors.initials: SA
authors.lastname: Wagner
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Petra
authors.initials: P
authors.lastname: Beli
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Rajat
authors.initials: R
authors.lastname: Gupta
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Trine A
authors.initials: TA
authors.lastname: Kristiansen
authors.affiliation: Division of Immune Cell Biology, MRC National Institute for Medical Research, Mill Hill, London, UK.
authors.forename: Dessislava
authors.initials: D
authors.lastname: Malinova
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Chiara
authors.initials: C
authors.lastname: Francavilla
authors.affiliation: Division of Immune Cell Biology, MRC National Institute for Medical Research, Mill Hill, London, UK.
authors.forename: Pavel
authors.initials: P
authors.lastname: Tolar
authors.affiliation: Department of Microbiology, Graduate Program in Immunology and Department of Internal Medicine, University of Iowa, Iowa City, IA, USA VAMC, Iowa City, IA, USA.
authors.forename: Gail A
authors.initials: GA
authors.lastname: Bishop
authors.affiliation: Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Hostager
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark chuna.choudhary@cpr.ku.dk.
authors.forename: Chunaram
authors.initials: C
authors.lastname: Choudhary
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis / Q000235:genetics*; D000074018:B-Cell CLL-Lymphoma 10 Protein; D001402:B-Lymphocytes / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D010766:Phosphorylation / Q000235:genetics; D040901:Proteomics; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000096:biosynthesis / Q000235:genetics*; D054875:Ubiquitination / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C584194:ANKRD13A protein, human; D000074018:B-Cell CLL-Lymphoma 10 Protein; C118591:BCL10 protein, human; D008565:Membrane Proteins; D016328:NF-kappa B; D011947:Receptors, Antigen, B-Cell; C558338:RNF31 protein, human; D044767:Ubiquitin-Protein Ligases
keywords: BCL10; BCR; RAB7A; phosphorylation; ubiquitylation