J Immunol 2002,
PMID: 12391233
Tcherkasowa, Anna Ewgenjewna; Adam-Klages, Sabine; Kruse, Marie-Luise; Wiegmann, Katja; Mathieu, Sabine; Kolanus, Waldemar; Krönke, Martin; Adam, Dieter
Factor associated with neutral sphingomyelinase activation (FAN) represents a p55 TNFR (TNF-R55)-associated protein essential for the activation of neutral sphingomyelinase. By means of the yeast interaction trap system, we have identified the scaffolding protein receptor for activated C-kinase (RACK)1 as an interaction partner of FAN. Mapping studies in yeast revealed that RACK1 is recruited to the C-terminal WD-repeat region of FAN and binds to FAN through a domain located within WD repeats V to VII of RACK1. Our data indicate that binding of both proteins is not mediated by linear motifs but requires folding into a secondary structure, such as the multibladed propeller characteristic of WD-repeat proteins. The interaction of FAN and RACK1 was verified in vitro by glutathione S-transferase-based coprecipitation assays as well as in eukaryotic cells by coimmunoprecipitation experiments. Colocalization studies in transfected cells suggest that TNF-R55 forms a complex with FAN and that this complex recruits RACK1 to the plasma membrane. Furthermore, activation of N-SMase by TNF was strongly enhanced when RACK1, FAN, and a noncytotoxic TNF-R55 mutant were expressed concurrently, suggesting RACK1 as a modulator of N-SMase activation. Together, these findings implicate RACK1 as a novel component of the signaling pathways of TNF-R55.
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Text Mining Data
N-SMase → TNF: "
Furthermore,
activation of
N-SMase by
TNF was strongly enhanced when RACK1, FAN, and a noncytotoxic TNF-R55 mutant were expressed concurrently, suggesting RACK1 as a modulator of N-SMase activation
"
N-SMase → RACK1: "
Furthermore, activation of N-SMase by TNF was strongly enhanced when RACK1 , FAN, and a noncytotoxic TNF-R55 mutant were expressed concurrently, suggesting RACK1 as a modulator of N-SMase activation
"
TNF → RACK1: "
Furthermore, activation of N-SMase by TNF was strongly enhanced when RACK1 , FAN, and a noncytotoxic TNF-R55 mutant were expressed concurrently, suggesting RACK1 as a modulator of N-SMase activation
"
Manually curated Databases
-
IRef Bind Interaction:
TNFRSF1A
—
NSMAF
-
IRef Bind Interaction:
TNFRSF1A
—
GNB2L1
-
IRef Bind Interaction:
GNB2L1
—
NSMAF
-
IRef Bind_translation Interaction:
TNFRSF1A
—
NSMAF
(coimmunoprecipitation)
-
IRef Bind_translation Interaction:
TNFRSF1A
—
GNB2L1
(coimmunoprecipitation)
-
IRef Bind_translation Interaction:
GNB2L1
—
NSMAF
(coimmunoprecipitation)
-
IRef Bind_translation Interaction:
GNB2L1
—
NSMAF
(affinity chromatography technology)
-
IRef Bind_translation Interaction:
GNB2L1
—
NSMAF
(two hybrid)
-
IRef Biogrid Interaction:
GNB2L1
—
NSMAF
(direct interaction, two hybrid)
-
IRef Biogrid Interaction:
GNB2L1
—
NSMAF
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
GNB2L1
—
NSMAF
(direct interaction, pull down)
-
IRef Hprd Interaction:
TNFRSF1A
—
GNB2L1
(in vitro)
-
IRef Hprd Interaction:
GNB2L1
—
NSMAF
(in vitro)
-
IRef Hprd Interaction:
GNB2L1
—
NSMAF
(in vivo)
-
IRef Hprd Interaction:
GNB2L1
—
NSMAF
(two hybrid)
-
IRef Ophid Interaction:
TNFRSF1A
—
GNB2L1
(aggregation, confirmational text mining)
-
NCI Pathway Database TNF receptor signaling pathway :
sTNF-alpha/TNFR1A/FAN/NSMASE complex (TNFRSF1A-NSMAF-SMPD2-TNF)
→
RACK1 (GNB2L1)
(modification, collaborate)
Evidence: mutant phenotype, assay, physical interaction
-
NCI Pathway Database TNF receptor signaling pathway :
sTNF-alpha/TNFR1A/FAN/NSMASE complex (TNFRSF1A-NSMAF-SMPD2-TNF)
→
sTNF-alpha/TNFR1A/FAN/NSMASE/RACK1 complex (TNFRSF1A-NSMAF-SMPD2-GNB2L1-TNF)
(modification, collaborate)
Evidence: mutant phenotype, assay, physical interaction
-
NCI Pathway Database TNF receptor signaling pathway :
RACK1 (GNB2L1)
→
sTNF-alpha/TNFR1A/FAN/NSMASE/RACK1 complex (TNFRSF1A-NSMAF-SMPD2-GNB2L1-TNF)
(modification, collaborate)
Evidence: mutant phenotype, assay, physical interaction
In total, 15 gene pairs are associated to this article in curated databases