EHD4 and CDH23 are interacting partners in cochlear hair cells

Soma Sengupta, Manju George, Katharine K. Miller, Khurram Naik, Jonathan Chou, Mary Ann Cheatham, Peter Dallos, Mayumi Naramura, Hamid Band, Jing Zheng*

*Corresponding author for this work

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Cadherin 23 (CDH23), a transmembrane protein localized near the tips of hair cell stereocilia in the mammalian inner ear, is important for delivering mechanical signals to the mechanoelectric transducer channels. To identify CDH23-interacting proteins, a membrane-based yeast two-hybrid screen of an outer hair cell (OHC) cDNA library was performed. EHD4, a member of the C-terminal EH domain containing a protein family involved in endocytic recycling, was identified as a potential interactor. To confirm the interaction, we first demonstrated the EHD4 mRNA expression in hair cells using in situ hybridization. Next, we showed that EHD4 co-localizes and co-immunoprecipitates with CDH23 in mammalian cells. Interestingly, the co-immunoprecipitation was found to be calcium-sensitive. To investigate the role of EHD4 in hearing, compound action potentials were measured in EHD4 knockout (KO) mice. Although EHD4 KO mice have normal hearing sensitivity, analysis of mouse cochlear lysates revealed a 2-fold increase in EHD1, but no increase in EHD2 or EHD3, in EHD4 KO cochleae compared with wild type, suggesting that a compensatory increase in EHD1 levels may account for the absence of a hearing defect in EHD4 KO mice. Taken together, these data indicate that EHD4 is a novel CDH23-interacting protein that could regulate CDH23 trafficking/localization in a calcium-sensitive manner.

Original languageEnglish (US)
Pages (from-to)20121-20129
Number of pages9
JournalJournal of Biological Chemistry
Volume284
Issue number30
DOIs
StatePublished - Jul 24 2009

Fingerprint

Auditory Hair Cells
Cadherins
Cells
Audition
Knockout Mice
Hearing
Cochlea
Outer Auditory Hair Cells
Proteins
Stereocilia
Calcium
Inner Ear
Transducers
Gene Library
Immunoprecipitation
Yeast
Action Potentials
Sensitivity analysis
In Situ Hybridization
Recycling

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sengupta, S., George, M., Miller, K. K., Naik, K., Chou, J., Cheatham, M. A., ... Zheng, J. (2009). EHD4 and CDH23 are interacting partners in cochlear hair cells. Journal of Biological Chemistry, 284(30), 20121-20129. https://doi.org/10.1074/jbc.M109.025668
Sengupta, Soma ; George, Manju ; Miller, Katharine K. ; Naik, Khurram ; Chou, Jonathan ; Cheatham, Mary Ann ; Dallos, Peter ; Naramura, Mayumi ; Band, Hamid ; Zheng, Jing. / EHD4 and CDH23 are interacting partners in cochlear hair cells. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 30. pp. 20121-20129.
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abstract = "Cadherin 23 (CDH23), a transmembrane protein localized near the tips of hair cell stereocilia in the mammalian inner ear, is important for delivering mechanical signals to the mechanoelectric transducer channels. To identify CDH23-interacting proteins, a membrane-based yeast two-hybrid screen of an outer hair cell (OHC) cDNA library was performed. EHD4, a member of the C-terminal EH domain containing a protein family involved in endocytic recycling, was identified as a potential interactor. To confirm the interaction, we first demonstrated the EHD4 mRNA expression in hair cells using in situ hybridization. Next, we showed that EHD4 co-localizes and co-immunoprecipitates with CDH23 in mammalian cells. Interestingly, the co-immunoprecipitation was found to be calcium-sensitive. To investigate the role of EHD4 in hearing, compound action potentials were measured in EHD4 knockout (KO) mice. Although EHD4 KO mice have normal hearing sensitivity, analysis of mouse cochlear lysates revealed a 2-fold increase in EHD1, but no increase in EHD2 or EHD3, in EHD4 KO cochleae compared with wild type, suggesting that a compensatory increase in EHD1 levels may account for the absence of a hearing defect in EHD4 KO mice. Taken together, these data indicate that EHD4 is a novel CDH23-interacting protein that could regulate CDH23 trafficking/localization in a calcium-sensitive manner.",
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Sengupta, S, George, M, Miller, KK, Naik, K, Chou, J, Cheatham, MA, Dallos, P, Naramura, M, Band, H & Zheng, J 2009, 'EHD4 and CDH23 are interacting partners in cochlear hair cells', Journal of Biological Chemistry, vol. 284, no. 30, pp. 20121-20129. https://doi.org/10.1074/jbc.M109.025668

EHD4 and CDH23 are interacting partners in cochlear hair cells. / Sengupta, Soma; George, Manju; Miller, Katharine K.; Naik, Khurram; Chou, Jonathan; Cheatham, Mary Ann; Dallos, Peter; Naramura, Mayumi; Band, Hamid; Zheng, Jing.

In: Journal of Biological Chemistry, Vol. 284, No. 30, 24.07.2009, p. 20121-20129.

Research output: Contribution to journalArticle

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AU - Sengupta, Soma

AU - George, Manju

AU - Miller, Katharine K.

AU - Naik, Khurram

AU - Chou, Jonathan

AU - Cheatham, Mary Ann

AU - Dallos, Peter

AU - Naramura, Mayumi

AU - Band, Hamid

AU - Zheng, Jing

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N2 - Cadherin 23 (CDH23), a transmembrane protein localized near the tips of hair cell stereocilia in the mammalian inner ear, is important for delivering mechanical signals to the mechanoelectric transducer channels. To identify CDH23-interacting proteins, a membrane-based yeast two-hybrid screen of an outer hair cell (OHC) cDNA library was performed. EHD4, a member of the C-terminal EH domain containing a protein family involved in endocytic recycling, was identified as a potential interactor. To confirm the interaction, we first demonstrated the EHD4 mRNA expression in hair cells using in situ hybridization. Next, we showed that EHD4 co-localizes and co-immunoprecipitates with CDH23 in mammalian cells. Interestingly, the co-immunoprecipitation was found to be calcium-sensitive. To investigate the role of EHD4 in hearing, compound action potentials were measured in EHD4 knockout (KO) mice. Although EHD4 KO mice have normal hearing sensitivity, analysis of mouse cochlear lysates revealed a 2-fold increase in EHD1, but no increase in EHD2 or EHD3, in EHD4 KO cochleae compared with wild type, suggesting that a compensatory increase in EHD1 levels may account for the absence of a hearing defect in EHD4 KO mice. Taken together, these data indicate that EHD4 is a novel CDH23-interacting protein that could regulate CDH23 trafficking/localization in a calcium-sensitive manner.

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Sengupta S, George M, Miller KK, Naik K, Chou J, Cheatham MA et al. EHD4 and CDH23 are interacting partners in cochlear hair cells. Journal of Biological Chemistry. 2009 Jul 24;284(30):20121-20129. https://doi.org/10.1074/jbc.M109.025668