A complex interaction pattern of CIS and SOCS2 with the leptin receptor

Lavens, Delphine; Montoye, Tony; Piessevaux, Julie; Zabeau, Lennart; Vandekerckhove, Joël; Gevaert, Kris; Becker, Walter; Eyckerman, Sven; Tavernier, Jan

doi:10.1242/jcs.02947

Cited by 51 publications

(48 citation statements)

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“…MfeI-XbaI digest of the first PCR fragment was ligated into an EcoRIXbaI opened pMG1-SVT vector (Eyckerman et al, 2001) with EcoRI site cloned C-terminal of the FLAG tag using site-directed mutagenesis with primers 59-GATGACGACGATAAGGAATTCTCGACCGTGGTAC-39 and 59-GTACCAC-GGTCGAGAATTCCTTATCGTCGTCATC-39. SacII-XbaI digest of the second PCR fragment was ligated into a SacII-XbaI opened pMET7-Etag-Cis vector (Lavens et al, 2006). Human RNF41 was cloned from the pMET7-Etag-hRNF41 vector into a pGEX-4T-2 vector using NotI-XbaI, downstream of glutathione Stransferase, generating the GST-RNF41 vector.…”

Section: Constructsmentioning

confidence: 99%

Reciprocal cross-regulation between RNF41 and USP8 controls cytokine receptor sorting and processing

Ceuninck

Wauman

Masschaele

et al. 2013

Journal of Cell Science

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SummaryThe mechanisms controlling the steady-state cell surface levels of cytokine receptors, and consequently the cellular response to cytokines, remain poorly understood. The number of surface-exposed receptors is a dynamic balance of de novo synthesis, transport to the plasma membrane, internalization, recycling, degradation and ectodomain shedding. We previously reported that the E3 ubiquitin ligase RING finger protein 41 (RNF41) inhibits basal lysosomal degradation and enhances ectodomain shedding of JAK2-associated cytokine receptors. Ubiquitin-specific protease 8 (USP8), an RNF41-interacting deubiquitylating enzyme (DUB) stabilizes RNF41 and is involved in trafficking of various transmembrane proteins. The present study identifies USP8 as a substrate of RNF41 and reveals that loss of USP8 explains the aforementioned RNF41 effects. RNF41 redistributes and ubiquitylates USP8, and reduces USP8 levels. In addition, USP8 knockdown functionally matches the effects of RNF41 ectopic expression on the model leptin and leukemia inhibitory factor (LIF) receptors. Moreover, RNF41 indirectly destabilizes the ESCRT-0 complex through suppression of USP8. Collectively, our findings demonstrate that RNF41 controls JAK2-associated cytokine receptor trafficking by acting as a key regulator of USP8 and ESCRT-0 stability. Balanced reciprocal cross-regulation of RNF41 and USP8 thus determines whether receptors are sorted for lysosomal degradation or recycling, this way regulating basal cytokine receptor levels.

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Section: Constructsmentioning

confidence: 99%

Reciprocal cross-regulation between RNF41 and USP8 controls cytokine receptor sorting and processing

Ceuninck

Wauman

Masschaele

et al. 2013

Journal of Cell Science

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“…As examples of known interaction partners of CIS and SOCS2, we used the intracellular receptor tyrosine motifs Tyr 402 of the EpoR (42) (47,48,52), is possible using the full-length LR. HEK293-T cells were cotransfected with the aforementioned baits and prey constructs encoding the different CIS and SOCS2 variants, combined with the STAT3-responsive rPAPI luciferase reporter.…”

Section: Cis Activity Depends On Elongin B/c Binding-to Investi-mentioning

confidence: 99%

Elongin B/C Recruitment Regulates Substrate Binding by CIS

Piessevaux

Ceuninck

Catteeuw

et al. 2008

Journal of Biological Chemistry

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SOCS proteins play a major role in the regulation of cytokine signaling. They are recruited to activated receptors and can suppress signaling by different mechanisms including targeting of the receptor complex for proteasomal degradation. The activity of SOCS proteins is regulated at different levels including transcriptional control and posttranslational modification. We describe here a novel regulatory mechanism for CIS, one of the members of this protein family. A CIS mutant deficient in recruitment of the Elongin B/C complex completely failed to suppress STAT5 activation. This deficiency was not caused by altered turnover of CIS but by loss of cytokine receptor interaction. Intriguingly, no such effect was seen for binding to MyD88. The interaction between CIS and the Elongin B/C complex, which depends on the levels of uncomplexed Elongin B/C, was easily disrupted. This regulatory mechanism may be unique for CIS, as similar mutations in SOCS1, -2, -3, -6, and -7 had no functional impact. Our findings indicate that the SOCS box not only plays a role in the formation of E3 ligase complexes but, at least for CIS, can also regulate the binding modus of SOCS boxcontaining proteins.Cytokines regulate multiple biological processes by activating specific cell surface receptor complexes. This leads to a series of signaling events, including activation of the Janus kinase (JAK) 2 /signal transducer and activator of transcription (STAT), phosphoinositol 3-kinase, phospholipase C␥, and mitogen-activated protein kinase pathways. The magnitude and duration of a cellular response is determined by the integration of different positive and negative signals. Mechanisms of signal attenuation are diverse and involve different protein families including phosphatases such as PTP-1B (protein tyrosine phosphatase 1B) and TCPTP (T-cell protein-tyrosine phosphatase) (1-5), members of the PIAS (protein inhibitors of activated STATs) (6 -8) and SOCS (suppressor of cytokine signaling) families (9 -11). SOCS proteins are induced by a broad range of extracellular ligands and function in a negative feedback loop to modulate signal transduction by multiple cytokine and growth factor receptors (12-14). The eight members of the SOCS family, SOCS1-7 and cytokine-inducible SH2-containing protein (CIS), share a common structure with a central SH2 domain, an amino-terminal domain of variable length and divergent sequence, and a carboxyl-terminal 40-amino acid module that is known as the SOCS box (9,15,16). The SH2 domain is the main determinant of target recognition by the SOCS proteins as it mediates interaction with phosphorylated tyrosine residues on their specific substrates (16 -18). In this way, SOCS proteins can suppress signaling by direct competition with signaling molecules for the phosphorylated recruitment sites. SOCS1 and SOCS3 can also inhibit JAK tyrosine kinase activity through their kinase inhibitory region, which is proposed to function as a pseudosubstrate blocking the catalytic cleft of the JAK kinase (19). Finally, SOCS prote...

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“…Remarkably, although SOCS2 does not bind the Tyr-985 motif, it can interfere with the binding of CIS to this site. This unexpected finding could be explained by the direct binding of SOCS2 on the CIS SOCS box, thereby targeting it for proteasomal degradation [6,10]. More detailed analysis using CIS mutants revealed that CIS binding to the LR and EpoR not only depends on its SH2 domain but also on its SOCS box, with a critical role for the CIS Tyr-253 and the Elongin B/C recruitment site.…”

Section: Lrmentioning

confidence: 99%

“…This way, protein interactions can be studied with the LR in its normal oligomeric configuration [6]. In another MAPPIT setup, we can use the wild-type EpoR ( Figure 1D) [7].…”

Section: Mappit (Mammalian Protein-protein Interaction Trap)mentioning

confidence: 99%

“…Since the EpoR signals via STAT5 and not STAT3 and since the levels of STAT5 are low in the commonly used HEK-293T cells [human embryonic kidney cells expressing the large T-antigen of SV40 (simian virus 40)], no reporter activation can be detected after stimulation with Epo (erythropoietin) allowing the use of the EpoR itself and mutants thereof as bait. To study the isolated tyrosine motifs of the LR, we replaced the LR cytosolic domain following the membraneproximal box1 motif (which is needed for the association of the JAK kinases) by flexible Gly-Gly-Ser repeats ( Figure 1E) [6]. To be able to use MAPPIT in haemopoietic cells that have no or very low STAT3 levels, βc (beta common) MAPPIT was created, which employs STAT5 as a signal transducer.…”

Section: Mappit (Mammalian Protein-protein Interaction Trap)mentioning

confidence: 99%

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MAPPIT: a versatile tool to study cytokine receptor signalling

Lemmens

Lievens

Tavernier

2008

Biochemical Society Transactions

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MAPPIT (mammalian protein-protein interaction trap) is a cytokine receptor-based two-hybrid method that operates in intact mammalian cells. A bait is fused C-terminally to a STAT (signal transducer and activator of transcription) recruitment-deficient receptor, whereas the prey is linked to functional STATbinding sites. When bait and prey interact a ligand-dependent complementation of the STAT recruitment deficiency occurs, leading to activation of a STAT-responsive reporter. MAPPIT is very well suited to study protein interactions involving activated cytokine receptors as the technique allows modification of the bait protein in a physiologically optimal environment. Cytokines and their receptorsCytokines are small messenger molecules that are involved in cell-cell communication and are critical to many vital processes such as embryonic development, growth, haemopoiesis and immune responses. They cannot cross the cellular membrane but make use of transmembrane receptors that capture cytokines and transmit their signal inside the cell. In this mini-review, we will mainly focus on homomeric type I cytokine receptors such as the EpoR (erythropoietin receptor), GHR (growth hormone receptor), LR (leptin receptor) and GCSFR (granulocyte colony-stimulating factor receptor). These cytokine receptors have no intrinsic kinase activity but make use of associated JAKs (Janus kinases). Upon ligand binding the JAKs are brought into close proximity enabling them to cross-phosphorylate each other. Subsequently the JAKs phosphorylate tyrosine residues in the cytoplasmic receptor tail, thereby creating docking sites for signalling molecules such as STAT (signal transducer and activator of transcription) proteins. After binding of these STATs they are phosphorylated in turn by the JAKs and translocate as dimers to the nucleus where they initiate transcription of target genes ( Figure 1A). Several mechanisms exist to modulate and terminate cytokine signalling. Among these, the SOCS (suppressor of cytokine signalling) protein Key words: cytokine receptor, mammalian protein-protein interaction trap (MAPPIT), signal transducer and activator of transcription (STAT), signal transduction, suppressor of cytokine signalling (SOCS), Toll-like receptor (TLR). Abbreviations used: βc, beta common; Epo, erythropoietin; EpoR, Epo receptor; GCSFR, granulocyte colony-stimulating factor receptor; GHR, growth hormone receptor; GM-CSF, granulocyte/macrophage colony-stimulating factor; gp130, glycoprotein 130; HEK-293T cells, human embryonic kidney cells expressing the large T-antigen of SV40 (simian virus 40); IRAK, interleukin-1-receptor-associated kinase; IRS4, insulin receptor substrate 4; JAK, Janus kinase; LR, leptin receptor; MAPPIT, mammalian protein-protein interaction trap; MyD88, myeloid differentiation primary response protein 88; Mal, MyD88 adaptor-like protein; PLCγ , phospholipase Cγ ; SH2, Src homology 2; CIS, cytokine-inducible SH2 domain-containing protein; SOCS, suppressor of cytokine signalling; STAT, signal transducer and ...

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A complex interaction pattern of CIS and SOCS2 with the leptin receptor

Cited by 51 publications

References 50 publications

Reciprocal cross-regulation between RNF41 and USP8 controls cytokine receptor sorting and processing

Reciprocal cross-regulation between RNF41 and USP8 controls cytokine receptor sorting and processing

Elongin B/C Recruitment Regulates Substrate Binding by CIS

MAPPIT: a versatile tool to study cytokine receptor signalling

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