TRAF2 Ser-11 Phosphorylation Promotes Cytosolic Translocation of the CD40 Complex To Regulate Downstream Signaling Pathways

Lauren M Workman; Laiqun Zhang; Yumei Fan; Weizhou Zhang; Hasem Habelhah

doi:10.1128/MCB.00429-19

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TRAF2 Ser-11 Phosphorylation Promotes Cytosolic Translocation of the CD40 Complex To Regulate Downstream Signaling Pathways

Journal article

Open access

Peer reviewed

TRAF2 Ser-11 Phosphorylation Promotes Cytosolic Translocation of the CD40 Complex To Regulate Downstream Signaling Pathways

Lauren M Workman, Laiqun Zhang, Yumei Fan, Weizhou Zhang and Hasem Habelhah

Molecular and cellular biology, Vol.40(9), e00429-19

04/13/2020

DOI: 10.1128/MCB.00429-19

PMCID: PMC7156217

PMID: 32041822

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7156217View

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Abstract

CD40 plays an important role in immune responses by activating the c-Jun N-terminal protein kinase (JNK) and NF-κB pathways; however, the precise mechanisms governing the spatiotemporal activation of these two signaling pathways are not fully understood. Here, using four different TRAF2-deficient cell lines (A20.2J, CH12.LX, HAP1, and mouse embryonic fibroblasts [MEFs]) reconstituted with wild-type or phosphorylation mutant forms of TRAF2, along with immunoprecipitation, immunoblotting, gene expression, and immunofluorescence analyses, we report that CD40 ligation elicits TANK-binding kinase 1 (TBK1)-mediated phosphorylation of TRAF2 at Ser-11. This phosphorylation interfered with the interaction between TRAF2's RING domain and membrane phospholipids and enabled translocation of the TRAF2 complex from CD40 to the cytoplasm. We also observed that this cytoplasmic translocation is required for full activation of the JNK pathway and the secondary phase of the NF-κB pathway. Moreover, we found that in the absence of Ser-11 phosphorylation, the TRAF2 RING domain interacts with phospholipids, leading to the translocation of the TRAF2 complex to lipid rafts, resulting in its degradation and activation of the noncanonical NF-κB pathway. Thus, our results provide new insights into the CD40 signaling mechanisms whereby Ser-11 phosphorylation controls RING domain-dependent subcellular localization of TRAF2 to modulate the spatiotemporal activation of the JNK and NF-κB pathways.

Phosphorylation

Animals

CD40 Antigens - metabolism

Cytoplasm - metabolism

Cytosol - metabolism

HEK293 Cells

Humans

JNK Mitogen-Activated Protein Kinases - metabolism

Mice

Mitogen-Activated Protein Kinases - metabolism

NF-kappa B - metabolism

Protein-Serine-Threonine Kinases - metabolism

Serine - metabolism

Signal Transduction - physiology

TNF Receptor-Associated Factor 2 - genetics

TNF Receptor-Associated Factor 2 - metabolism

TNF Receptor-Associated Factor 6 - metabolism

Transcription Factor RelA - metabolism

Details

Title: Subtitle: TRAF2 Ser-11 Phosphorylation Promotes Cytosolic Translocation of the CD40 Complex To Regulate Downstream Signaling Pathways
Creators: Lauren M Workman - University of Iowa
Laiqun Zhang - University of Iowa
Yumei Fan - Hebei Normal University
Weizhou Zhang - University of Iowa
Hasem Habelhah - University of Iowa
Resource Type: Journal article
Publication Details: Molecular and cellular biology, Vol.40(9), e00429-19
DOI: 10.1128/MCB.00429-19
PMID: 32041822
PMCID: PMC7156217
ISSN: 0270-7306
eISSN: 1098-5549
Grant note: P30 CA086862 / NCI NIH HHS
Language: English
Date published: 04/13/2020
Academic Unit: Pathology; Radiation Oncology
Record Identifier: 9984186580402771

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