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

Reo Maeda, Katie S. Kindt, Weike Mo, Clive P. Morgan, Timothy Erickson, Hongyu Zhao, Rachel Clemens-Grisham, Peter Barr-Gillespie, Teresa Nicolson

Research output: Contribution to journalArticle

67 Citations (Scopus)

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 channellike 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 determinewhether 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.

Original languageEnglish (US)
Pages (from-to)12907-12912
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number35
DOIs
StatePublished - Sep 2 2014

Fingerprint

Zebrafish
Two-Hybrid System Techniques
Protein Isoforms
Auditory Hair Cells
Vestibular Hair Cells
Proteins
Fishes
Membranes
link protein

ASJC Scopus subject areas

  • General

Cite this

Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2. / Maeda, Reo; Kindt, Katie S.; Mo, Weike; Morgan, Clive P.; Erickson, Timothy; Zhao, Hongyu; Clemens-Grisham, Rachel; Barr-Gillespie, Peter; Nicolson, Teresa.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, No. 35, 02.09.2014, p. 12907-12912.

Research output: Contribution to journalArticle

Maeda, Reo ; Kindt, Katie S. ; Mo, Weike ; Morgan, Clive P. ; Erickson, Timothy ; Zhao, Hongyu ; Clemens-Grisham, Rachel ; Barr-Gillespie, Peter ; Nicolson, Teresa. / Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2. In: Proceedings of the National Academy of Sciences of the United States of America. 2014 ; Vol. 111, No. 35. pp. 12907-12912.
@article{275ec85a14ca4bc4ab14d15df6110250,
title = "Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2",
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 channellike 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 determinewhether 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.",
author = "Reo Maeda and Kindt, {Katie S.} and Weike Mo and Morgan, {Clive P.} and Timothy Erickson and Hongyu Zhao and Rachel Clemens-Grisham and Peter Barr-Gillespie and Teresa Nicolson",
year = "2014",
month = "9",
day = "2",
doi = "10.1073/pnas.1402152111",
language = "English (US)",
volume = "111",
pages = "12907--12912",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "35",

}

TY - JOUR

T1 - Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2

AU - Maeda, Reo

AU - Kindt, Katie S.

AU - Mo, Weike

AU - Morgan, Clive P.

AU - Erickson, Timothy

AU - Zhao, Hongyu

AU - Clemens-Grisham, Rachel

AU - Barr-Gillespie, Peter

AU - Nicolson, Teresa

PY - 2014/9/2

Y1 - 2014/9/2

N2 - 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 channellike 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 determinewhether 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.

AB - 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 channellike 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 determinewhether 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.

UR - http://www.scopus.com/inward/record.url?scp=84907228023&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84907228023&partnerID=8YFLogxK

U2 - 10.1073/pnas.1402152111

DO - 10.1073/pnas.1402152111

M3 - Article

C2 - 25114259

AN - SCOPUS:84907228023

VL - 111

SP - 12907

EP - 12912

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 35

ER -