The E3 Ubiquitin Ligase Atrophin Interacting Protein 4 Binds Directly To The Chemokine Receptor CXCR4 Via a Novel WW Domain-mediated Interaction

Bhandari, Deepali; Robia, Seth L.; Marchese, Adriano

doi:10.1091/mbc.e08-03-0308

Cited by 80 publications

(136 citation statements)

References 45 publications

Supporting

Mentioning

131

Contrasting

Order By: Relevance

“…Most Nedd4-2-targeted proteins interact with its WW3-4 domains. Interestingly, our study revealed that the N-terminal region of TRPV6 interacts with WW1 and WW2 domains, a situation similar to the interaction between CXCR4 and AIP4 (44). However, the interaction between WW1-2 domains and TRPV6 is not indispensable for the association between Nedd4-2 and TRPV6, because other domains, such as the HECT and C2 domains, significantly interact with the TRPV6 C-terminal region.…”

Section: Discussionmentioning

confidence: 48%

“…However, not every protein directly binds with the WW domain of Nedd4-type ubiquitin E3 ligases through a PY motif. A chemokine receptor, CXCR4, which has no PY motif, binds with the WW1 or WW2 domains of E3 ubiquitin ligase AIP4 via a 324/ 325 double serine in its C-terminal region (44). PTEN even interacts with the C2 and HECT domains of Nedd4 (45).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Down-regulation of Intestinal Apical Calcium Entry Channel TRPV6 by Ubiquitin E3 Ligase Nedd4-2

Zhang¹,

Na²,

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Nedd4-2 is an archetypal HECT ubiquitin E3 ligase that disposes target proteins for degradation. Because of the proven roles of Nedd4-2 in degradation of membrane proteins, such as epithelial Na ؉ channel, we examined the effect of Nedd4-2 on the apical Ca 2؉ channel TRPV6, which is involved in transcellular Ca 2؉ transport in the intestine using the Xenopus laevis oocyte system. We demonstrated that a significant amount of Nedd4-2 protein was distributed to the absorptive epithelial cells in ileum, cecum, and colon along with TRPV6. When coexpressed in oocytes, Nedd4-2 and, to a lesser extent, Nedd4 down-regulated the protein abundance and Ca 2؉ influx of TRPV6 and TRPV5, respectively. TRPV6 ubiquitination was increased, and its stability was decreased by Nedd4-2. The Nedd4-2 inhibitory effects on TRPV6 were partially blocked by proteasome inhibitor MG132 but not by the lysosome inhibitor chloroquine. The rate of TRPV6 internalization was not significantly altered by Nedd4-2. The HECT domain was essential to the inhibitory effect of Nedd4-2 on TRPV6 and to their association. The WW1 and WW2 domains interacted with TRPV6 terminal regions, and a disruption of the interactions by D204H and D376H mutations in the WW1 and WW2 domains increased TRPV6 ubiquitination and degradation. Thus, WW1 and WW2 may serve as a molecular switch to limit the ubiquitination of TRPV6 by the HECT domain. In conclusion, Nedd4-2 may regulate TRPV6 protein abundance in intestinal epithelia by controlling TRPV6 ubiquitination.

show abstract

Section: Discussionmentioning

confidence: 48%

Section: Discussionmentioning

confidence: 99%

Down-regulation of Intestinal Apical Calcium Entry Channel TRPV6 by Ubiquitin E3 Ligase Nedd4-2

Zhang¹,

Na²,

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…The modulation of the p63-Itch binding by phosphorylation could be a mechanism more widespread and regulate also the interaction of Itch-E3 ligase with other proteins, such as observed in the interaction of Itch with the chemokine receptor CXCR4. 111 This last one occurs through WW-Itch domains and a phosphorylated serine residue of a recognition sequence that does not contain proline residues. 111 The scheme in Figure 7 can be generalized for other PY interactions with WW domains, as the PY motif of p63, is also present in other proteins including for example RASSF5 where the PPxY motif interact with the WW domain of Itch 112 in a manner very similar to p63.…”

Section: Discussionmentioning

confidence: 99%

“…111 This last one occurs through WW-Itch domains and a phosphorylated serine residue of a recognition sequence that does not contain proline residues. 111 The scheme in Figure 7 can be generalized for other PY interactions with WW domains, as the PY motif of p63, is also present in other proteins including for example RASSF5 where the PPxY motif interact with the WW domain of Itch 112 in a manner very similar to p63. All these proteins are characterized by the nearby presence of a (T/S)P motif, which is a potential recognition site of the WW domain of the IV group present in the prolyl-isomerase Pin1.…”

Section: Discussionmentioning

confidence: 99%

p63 threonine phosphorylation signals the interaction with the WW domain of the E3 ligase Itch

et al. 2014

View full text Add to dashboard Cite

Both in epithelial development as well as in epithelial cancers, the p53 family member p63 plays a crucial role acting as a master transcriptional regulator. P63 steady state protein levels are regulated by the E3 ubiquitin ligase Itch, via a physical interaction between the PPxY consensus sequence (PY motif) of p63 and one of the 4 WW domains of Itch; this substrate recognition process leads to protein-ubiquitylation and p63 proteasomal degradation. The interaction of the WW domains, a highly compact protein-protein binding module, with the short proline-rich sequences is therefore a crucial regulatory event that may offer innovative potential therapeutic opportunity. Previous molecular studies on the Itch-p63 recognition have been performed in vitro using the Itch-WW2 domain and the peptide interacting fragment of p63 (pep63), which includes the PY motif. Itch-WW2-pep63 interaction is also stabilized in vitro by the conformational constriction of the S-S cyclization in the p63 peptide. The PY motif of p63, as also for other proteins, is characterized by the nearby presence of a (T/S)P motif, which is a potential recognition site of the WW domain of the IV group present in the prolyl-isomerase Pin1. In this study, we demonstrate, by in silico and spectroscopical studies using both the linear pep63 and its cyclic form, that the threonine phosphorylation of the (T/S)PPPxY motif may represent a crucial regulatory event of the Itch-mediated p63 ubiquitylation, increasing the Itch-WW domains-p63 recognition event and stabilizing in vivo the Itch-WW-p63 complex. Moreover, our studies confirm that the subsequently trans/cis proline isomerization of (T/S)P motif by the Pin1 prolyl-isomerase, could modulate the E3-ligase interaction, and that the (T/S) p P trans PPxY motif represent the best conformer for the ItchWW-(T/S)PPPxY motif recognition.

show abstract

“…CXCR4 mediated chemotaxis along a gradient of its cognate ligand, stromal-derived factor 1␣ (SDF-1␣/CXCL12), and supports metastasis of cancer to the bone marrow (25,28). Ligand-induced lysosomal degradation of CXCR4 is critical for downstream signal attenuation and is mediated by the HECT family E3 ligase atrophininteracting protein 4 (AIP4/Itch) (29,30) and deubiquitinase ubiquitin-specific protease 14 (USP14) (31). CXCR4 activation has also been linked to ubiquitination of Hrs by AIP4, and its trafficking proceeds along an Hrs-dependent pathway (32), suggesting that alteration in the Hrs ubiquitination status may modulate CXCR4 turnover.…”

mentioning

confidence: 99%

The Deubiquitinating Enzyme USP8 Promotes Trafficking and Degradation of the Chemokine Receptor 4 at the Sorting Endosome

Berlin¹,

Higginbotham

Dise

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Reversible ubiquitination orchestrated by the opposition of ubiquitin ligases and deubiquitinating enzymes mediates endocytic trafficking of cell surface receptors for lysosomal degradation. Ubiquitin-specific protease 8 (USP8) has previously been implicated in endocytosis of several receptors by virtue of their deubiquitination. The present study explores an indirect role for USP8 in cargo trafficking through its regulation of the chemokine receptor 4 (CXCR4). Contrary to the effects of USP8 loss on enhanced green fluorescent protein, we find that USP8 depletion stabilizes CXCR4 on the cell surface and attenuates receptor degradation without affecting its ubiquitination status. In the presence of ligand, diminished CXCR4 turnover is accompanied by receptor accumulation on enlarged early endosomes and leads to enhancement of phospho-ERK signaling. Perturbation in CXCR4 trafficking, resulting from USP8 inactivation, occurs at the ESCRT-0 checkpoint, and catalytic mutation of USP8 specifically targeted to the ESCRT-0 complex impairs the spatial and temporal organization of the sorting endosome. USP8 functionally opposes the ubiquitin ligase AIP4 with respect to ESCRT-0 ubiquitination, thereby promoting trafficking of CXCR4. Collectively, our findings demonstrate a functional cooperation between USP8, AIP4, and the ESCRT-0 machinery at the early sorting phase of CXCR4 and underscore the versatility of USP8 in shaping trafficking events at the earlyto-late endosome transition.Endocytosis and trafficking of cell surface receptors is essential for organized signal transduction and maintenance of homeostasis. Malfunctions along the molecular pathways governing endocytosis can lead to a wide range of human pathologies including metabolic disorders, autoimmune diseases, and cancer (1, 2). Ubiquitination, a reversible post-translational modification of proteins (3), mediates spatial and temporal aspects of endocytosis by dictating macromolecular complex assembly and cargo fate (4). Although polyubiquitination linked through Lys 48 of ubiquitin targets substrates for proteasomal degradation (5), mono-and Lys 63 -linked ubiquitination signal cargo trafficking (6), and modulate function of endocytic sorting machinery (7). The reversible nature of ubiquitination is imparted by deubiquitinating enzymes (DUBs) 4 and enables dynamic regulation of these ubiquitin-dependent processes in the cell (8).Trafficking of endocytosed cargo for degradation by the lysosome occurs in a series of discrete selection stages that utilize the endosomal sorting complexes required for transport (ESCRTs) -0, -I, -II, and -III. The ESCRT-0 complex, consisting of the adaptor proteins hepatocyte growth factor-regulated substrate (Hrs) and signal transducing adaptor molecule (STAM), resides at the sorting endosome and functions in the first step of cargo selection. ESCRT-0 partitions the endocytosed material arriving on the early endosomes between recycling and the multivesicular body (9 -11) and interacts with downstream ESCRT machinery (12, 13) respon...

show abstract

The E3 Ubiquitin Ligase Atrophin Interacting Protein 4 Binds Directly To The Chemokine Receptor CXCR4 Via a Novel WW Domain-mediated Interaction

Cited by 80 publications

References 45 publications

Down-regulation of Intestinal Apical Calcium Entry Channel TRPV6 by Ubiquitin E3 Ligase Nedd4-2

Down-regulation of Intestinal Apical Calcium Entry Channel TRPV6 by Ubiquitin E3 Ligase Nedd4-2

p63 threonine phosphorylation signals the interaction with the WW domain of the E3 ligase Itch

The Deubiquitinating Enzyme USP8 Promotes Trafficking and Degradation of the Chemokine Receptor 4 at the Sorting Endosome

Contact Info

Product

Resources

About