title: PDZ domain from Dishevelled -- a specificity study.
abstract: Intracellular signaling cascades induced by Wnt proteins play a key role in developmental processes and are implicated in cancerogenesis. It is still unclear how the cell determines which of the three possible Wnt response mechanisms should be activated, but the decision process is most likely dependent on Dishevelled proteins. Dishevelled family members interact with many diverse targets, however, molecular mechanisms underlying these binding events have not been comprehensively described so far. Here, we investigated the specificity of the PDZ domain from human Dishevelled-2 using C-terminal phage display, which led us to identification of a leucine-rich binding motif strongly resembling the consensus sequence of a nuclear export signal. PDZ interactions with several peptide and protein motifs (including the nuclear export signal sequence from Dishevelled-2 protein) were investigated in detail using fluorescence spectroscopy, mutational analysis and immunoenzymatic assays. The experiments showed that the PDZ domain can bind the nuclear export signal sequence of the Dishevelled-2 protein. Since the intracellular localization of Dishevelled is governed by nuclear localization and nuclear export signal sequences, it is possible that the intramolecular interaction between PDZ domain and the export signal could modulate the balance between nuclear and cytoplasmic pool of the Dishevelled protein. Such a regulatory mechanism would be of utmost importance for the differential activation of Wnt signaling cascades, leading to selective promotion of the nucleus-dependent Wnt β-catenin pathway at the expense of non-canonical Wnt signaling.
authors.affiliation: Faculty of Biotechnology, Department of Protein Engineering, University of Wrocław, Wrocław, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Śmietana
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Mateja
authors.forename: Artur
authors.initials: A
authors.lastname: Krężel
authors.forename: Jacek
authors.initials: J
authors.lastname: Otlewski
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry*; D000595:Amino Acid Sequence; D000072261:Dishevelled Proteins; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D054731:PDZ Domains; D019151:Peptide Library; D010750:Phosphoproteins / Q000737:chemistry*; D011485:Protein Binding; D015398:Signal Transduction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C000606581:DVL2 protein, human; D000072261:Dishevelled Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009842:Oligopeptides; D019151:Peptide Library; D010750:Phosphoproteins