PUBLICATION

Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2

Authors
Maeda, R., Kindt, K.S., Mo, W., Morgan, C.P., Erickson, T., Zhao, H., Clemens-Grisham, R., Barr-Gillespie, P.G., Nicolson, T.
ID
ZDB-PUB-140813-4
Date
2014
Source
Proceedings of the National Academy of Sciences of the United States of America   111(35): 12907-12 (Journal)
Registered Authors
Clemens-Grisham, Rachel, Erickson, Timothy, Kindt, Katie, Maeda, Reo, Mo, Weike, Nicolson, Teresa
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Cadherins/genetics
  • Cadherins/metabolism*
  • Evolution, Molecular
  • Gene Expression Regulation, Developmental
  • HEK293 Cells
  • Hair Cells, Auditory/physiology*
  • Hair Cells, Vestibular/physiology
  • Humans
  • Mechanotransduction, Cellular/genetics
  • Mechanotransduction, Cellular/physiology*
  • Membrane Proteins/genetics
  • Membrane Proteins/metabolism*
  • Mice
  • Phylogeny
  • Protein Precursors/genetics
  • Protein Precursors/metabolism
  • Two-Hybrid System Techniques
  • Zebrafish/genetics
  • Zebrafish/growth & development
  • Zebrafish/physiology*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed
25114259 Full text @ Proc. Natl. Acad. Sci. USA
Abstract
The tip link protein protocadherin 15 (PCDH15) is a central component of the mechanotransduction complex in auditory and vestibular hair cells. PCDH15 is hypothesized to relay external forces to the mechanically gated channel located near its cytoplasmic C terminus. How PCDH15 is coupled to the transduction machinery is not clear. Using a membrane-based two-hybrid screen to identify proteins that bind to PCDH15, we detected an interaction between zebrafish Pcdh15a and an N-terminal fragment of transmembrane channel-like 2a (Tmc2a). Tmc2a is an ortholog of mammalian TMC2, which along with TMC1 has been implicated in mechanotransduction in mammalian hair cells. Using the above-mentioned two-hybrid assay, we found that zebrafish Tmc1 and Tmc2a can interact with the CD1 or CD3 cytoplasmic domain isoforms of Pcdh15a, and this interaction depends on the common region shared between the two Pcdh15 isoforms. Moreover, an interaction between mouse PCDH15-CD3 and TMC1 or TMC2 was observed in both yeast two-hybrid assays and coimmunoprecipitation experiments. To determine whether the Pcdh15-Tmc interaction is relevant to mechanotransduction in vivo, we overexpressed N-terminal fragments of Tmc2a in zebrafish hair cells. Overexpression of the Tmc2a N terminus results in mislocalization of Pcdh15a within hair bundles, together with a significant decrease in mechanosensitive responses, suggesting that a Pcdh15a-Tmc complex is critical for mechanotransduction. Together, these results identify an evolutionarily conserved association between the fish and mouse orthologs of PCDH15 and TMC1 and TMC2, supporting the notion that TMCs are key components of the transduction complex in hair cells.
Genes / Markers
Figures
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Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes