ZFIN ID: ZDB-PUB-080826-12
Identification and functional characterisation of an N-terminal oligomerisation domain for polycystin-2
Feng, S., Okenka, G.M., Bai, C.X., Streets, A.J., Newby, L.J., Dechant, B.T., Tsiokas, L., Obara, T., and Ong, A.C.
Date: 2008
Source: The Journal of biological chemistry   283(42): 28471-28479 (Journal)
Registered Authors: Obara, Tomoko
Keywords: none
MeSH Terms:
  • Animals
  • Dimerization
  • Electrophysiology/methods
  • Humans
  • Immunohistochemistry/methods
  • Models, Biological
  • Mutation
  • Plasmids/metabolism
  • Polycystic Kidney Diseases/metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • TRPP Cation Channels/chemistry*
  • TRPP Cation Channels/metabolism
  • Two-Hybrid System Techniques
  • Zebrafish
PubMed: 18701462 Full text @ J. Biol. Chem.
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ABSTRACT
Autosomal dominant polycystic kidney disease (ADPKD), the most common inherited cause of kidney failure, is caused by mutations in either PKD1 (85%) or PKD2 (15%). The PKD2 protein, polycystin-2 (PC2 or TRPP2), is a member of the transient receptor potential (TRP) superfamily and functions as a non-selective calcium channel. PC2 has been found to form oligomers in native tissues suggesting that it may form functional homo- or heterotetramers with other subunits, similar to other TRP channels. Our experiments unexpectedly revealed that PC2 mutant proteins lacking the known C-terminal dimerisation domain, were still able to form oligomers and co-immunoprecipitate full-length PC2, implying the possible existence of a proximal dimerisation domain. Using yeast two-hybrid and biochemical assays, we have mapped an alternative dimerisation domain to the N-terminus of PC2 (NT2 1-223, L224X). Functional characterisation of this domain demonstrated that it was sufficient to induce cyst formation in zebrafish embryos and inhibit PC2 surface currents in mIMCD3 cells probably by a dominant-negative mechanism. In summary, we propose a model for PC2 assembly as a functional tetramer which depends on both C- and N-terminal dimerisation domains. These results have significant implications for our understanding of PC2 function and disease pathogenesis in ADPKD and provide a new strategy for studying PC2 function.
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