The Eukaryotic Linear Motif resource for
Functional Sites in Proteins
Accession:
Functional site class:
Wnt-Wingless palmitoylation site
Functional site description:
Glycosylation of Wnt proteins in the Endoplasmic Reticulum is required for their proper function but is in competition with disulphide bond formation. Porcupine is required for correct processing of Wnt in the ER and porc mutants do not secrete Wingless. It appears to achieve this by palmitoylating a conserved Cys residue, thereby interfering with the disulphide bond formation process and allowing glycosylation to complete.
ELM Description:
Palmitoylation site on Wnt recognised by Porcupine (Tanaka,2002, Willert,2003). The site is only approximately localised (23 residue region 83-106 in Drosophila Wingless P09615) and the residues actually recognised by Porcupine are not yet known. Disulphide bond formation competes with the N-glycosylation required for Wnt function. Palmitoylation of a conserved Cys residue delays disulphide bond formation and promotes glycosylation. This regular expression pattern spans the palmitoylated Cys and the nearby glycosylation site. It finds typical Wnts but will miss some of the more divergent Wnt paralogues.
Pattern: [ETA](C)[QERK]..F...RWNC[ST]
Pattern Probability: 2.518e-13
Present in taxons: Drosophila melanogaster Homo sapiens Metazoa Xenopus laevis
Interaction Domain:
MBOAT (PF03062) MBOAT family (Stochiometry: 1 : 1)
o See 1 Instance for MOD_WntLipid
o Abstract
The protein Wingless (Wg) is the archetype of the Wnt protein family (Cadigan,1997, Polakis,2000). Drosophila has several Wnt paralogues and vertebrates have large Wnt multigene families. Wnt proteins are secreted developmental regulators that bind to cell surface receptor complexes of the Frizzled family, LRP5/6 and Kremen1/2 (Pandur,2001, Mao,2002). Intracellular signalling by Frizzled receptors mainly occurs through the beta-catenin pathway. Wnt is glycosylated at conserved sites (Blasband,1992), which is absolutely required for binding to its receptor. Processing of Wnt in the Endoplasmic Reticulum is unusually slow so that secretion is "inefficient". The Drosophila protein Porcupine influences Wingless processing in the ER, so that porc mutant cells retain Wingless (Kadowaki,1996). A clue to the role of Porcupine was provided by the finding that it belongs to a family of transmembrane acyltransferase enzymes (Hofmann,2000). It has now been shown by mass spectrometry that secreted mouse Wnt3a and Drosophila Wnt8, when overexpressed, are partially palmitoylated on a conserved Cys residue: C77 and C51, respectively (Willert,2003). The region of interaction with Porcupine has been localised to a 23-residue peptide segment spanning this residue and Porcupine strongly facilitates Wnt glycosylation (Tanaka,2002). It is reasonable to infer that Porcupine is the acyltransferase.
Why is Wnt palmitoylated? One of the conserved N-glycosylation sites (MOD_N-GLC_1 and MOD_N-GLC_2) has the sequence NCS, while the second conserved site is CNxS, where the conserved Cys residues will form disulphide bonds, quite possibly with each other. Glycosylation occurs on exposed peptide segments, hence a disulphide-linked moiety will be unavailable for glycosylation. For Wnts, there must be a mechanism to prevent disulphide formation until glycosylation is complete. The palmitoylated Cys precedes the NCS glycosylated Asn by 11 residues. Palmitoylation will have two effects: (i) preventing Wnt from completing folding into the native structure, and (ii) targeting Wnt to the ER membrane where glycosyltransferase enzymes reside. After Wnt glycosylation, further processing should include deacylation to allow disulphide formation and folding to complete. Secretion of palmitoylated Wnt may be an artefact of overexpression, overloading the processing machinery. Mass spectrometry should then reveal that fully processed natural Wnt is not palmitoylated. Candidates for the deacylase may be the Palmitoyl protein thioesterases (PPT1/2), depalmitoylating enzymes found predomininantly in lysosomes but also in other compartments such as neuronal synaptosomes (Ahtiainen,2003). It cannot yet be excluded that secreted palmitoylated Wnt is indeed functional. Weak, indirect evidence is suggested by domains (SM00192; PF00057) found in LRP5/6 that are also found in LDL receptors, while the Dickkopf family of Wingless inhibitors have a domain that is related to colipases (Aravind,1998, Hofmann,2000), however current evidence indicates that Dickkopf proteins do not physically interact with Wnt itself. This emerging and tentative scheme needs further testing. The whole system seems unnecessarily complicated, unless it is part of a regulated mechanism used to modulate levels of Wnt secretion during development. Most palmitoylated proteins are intracellular but a few other extracellular proteins are found to be palmitoylated, including glypicans (Edgren,1997), surfactant protein C (ten Brinke,2001) and Apolipoprotein B (Zhao,2000). Exported signalling proteins of the hedgehog family are also palmitoylated (Pepinsky,1998, Chamoun,2001).
o 13 selected references:

o 4 GO-Terms:

o 1 Instance for MOD_WntLipid
(click table headers for sorting; Notes column: =Number of Switches, =Number of Interactions)
Acc., Gene-, NameStartEndSubsequenceLogic#Ev.OrganismNotes
P27467 Wnt3a
WNT3A_MOUSE
76 89 GIQECQHQFRGRRWNCTTVS TP 3 Mus musculus (House mouse)
Please cite: ELM 2016-data update and new functionality of the eukaryotic linear motif resource. (PMID:26615199)

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