Related Experiment Video
Updated: May 9, 2026

Analysis of SCAP N-glycosylation and Trafficking in Human Cells
Published on: November 8, 2016
Sphingosine-1-phosphate is a missing cofactor for the E3 ubiquitin ligase TRAF2
Sergio E Alvarez1, Kuzhuvelil B Harikumar, Nitai C Hait
1Department of Biochemistry and Molecular Biology and the Massey Cancer Center, Virginia Commonwealth University School of Medicine, 1101 E. Marshall Street, Richmond, Virginia 23298, USA.
Sphingosine-1-phosphate (S1P) acts as a cofactor for TRAF2, a key enzyme in TNF-alpha signaling. This discovery reveals a new mechanism for regulating NF-kappaB activation, crucial for inflammation and immunity.
Area of Science:
- Cellular signaling pathways
- Molecular biology
- Biochemistry
Background:
- Tumor necrosis factor (TNF)-alpha signaling activates NF-kappaB, a transcription factor vital for immune responses.
- TRAF2 (TNF receptor-associated factor 2) is essential for NF-kappaB activation but its direct role in RIP1 ubiquitination was unclear.
- Sphingosine kinase 1 (SphK1) produces sphingosine-1-phosphate (S1P), a lipid mediator involved in cell survival.
Purpose of the Study:
- To investigate the role of S1P in TRAF2-mediated NF-kappaB activation.
- To determine if S1P acts as a cofactor for TRAF2's E3 ligase activity.
- To elucidate the mechanism of TRAF2-mediated RIP1 ubiquitination.
Main Methods:
- In vitro ubiquitination assays using recombinant TRAF2 and RIP1.
- Measurement of S1P production and its effect on NF-kappaB pathway components.
- Analysis of S1P binding to TRAF2.
Main Results:
- SphK1 and intracellular S1P are necessary for RIP1 ubiquitination, IKK/IkappaBalpha phosphorylation, and NF-kappaB activation.
- S1P directly binds to TRAF2's RING domain and enhances its E3 ligase activity.
- S1P specifically promotes lysine-63-linked polyubiquitination of RIP1 by TRAF2.
Conclusions:
- TRAF2 is a novel intracellular target of S1P.
- S1P is a crucial cofactor for TRAF2 E3 ubiquitin ligase activity, regulating lysine-63-linked polyubiquitination.
- This finding establishes a new paradigm for NF-kappaB pathway regulation by SphK1/S1P and TRAF2.
Related Concept Videos
Regulation of Nuclear Protein Sorting
Phosphoinositides and PIPs
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
Coat Assembly and GTPases
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
Cotranslational Protein Translocation
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Tail-anchoring of Proteins in the ER Membrane
Membrane Asymmetry Regulating Transporters
Flippase
Eukaryotic flippases are type-IV P-type ATPases or P4-ATPases belonging to P-type ATPase family proteins that are membrane-bound pumps involved in the ATP-mediated transport of ions and molecules across the membrane. Flippases flip specific phospholipids from the outer to the inner leaflet of a membrane. All P4-ATPases have one...

