| 2014 |
TMIE forms a ternary complex with the tip-link component PCDH15 and its binding partner TMHS/LHFPL5 in cochlear hair cells. Alternative splicing of the PCDH15 cytoplasmic domain regulates formation of this ternary complex. Homozygous Tmie-null mutation abolishes transducer currents, and subtle Tmie mutations that disrupt interactions between TMIE and tip links impair transduction, establishing TMIE as an essential component of the mechanotransduction machinery that functionally couples the tip link to the transduction channel. |
Co-immunoprecipitation, pulldown assays, electrophysiology in Tmie-null and point-mutant mice, alternative splicing analysis |
Neuron |
High |
25467981
|
| 2020 |
TMIE binds TMC1/2 and is required for TMC1/2 to form functional mechanotransduction channels; a TMIE mutation that perturbs TMC1/2 binding abolishes mechanotransduction. N-terminal TMIE deletions alter channel responses to mechanical force. The C-terminal cytoplasmic domain of TMIE contains charged residues that mediate binding to phospholipids including PIP2; deafness-linked C-terminal point mutations disrupt phospholipid binding, sensitize the channel to PIP2 depletion, and alter unitary conductance and ion selectivity, defining TMIE as a channel subunit with a phospholipid-sensing domain. |
Co-immunoprecipitation (TMIE–TMC1/2 binding), site-directed mutagenesis, whole-cell and single-channel electrophysiology in hair cells, PIP2 depletion assays, N-terminal deletion constructs |
Neuron |
High |
32343945
|
| 2019 |
In zebrafish tmie mutants, GFP-tagged Tmc1 and Tmc2b fail to target to the hair bundle, while Tmie overexpression strongly enhances Tmc1/Tmc2b targeting to stereocilia. Systematic deletion/replacement of Tmie peptide segments showed that the extracellular region and transmembrane domain are required for both mechanosensitivity and Tmc2b-GFP bundle expression, indicating TMIE's role is to target and stabilize Tmc channel subunits at the site of mechanotransduction. |
Fluorescence imaging of GFP-tagged TMC subunits in tmie zebrafish mutants, domain-deletion and chimeric Tmie rescue constructs, functional electrophysiology |
PLoS genetics |
High |
30726219
|
| 2025 |
Mouse TMC1/2 with a Fyn lipidation tag can reach the cell surface and form mechanosensitive channels in heterologous cells without TMIE, but TMIE robustly stimulates TMC1/2 channel activity by modulating their gating. Palmitoylation sites C76/C77 on TMIE are essential for this stimulation; mutating these sites eliminates TMIE's ability to enhance TMC1/2 gating. TMC1+TMIE and TMC2+TMIE form 18 pS and 24 pS single channels, respectively, with biophysical properties similar to the native mechanotransduction channel. |
Heterologous expression of Fyn-tagged TMC1/2 ± TMIE in non-hair cells, single-channel patch-clamp electrophysiology, site-directed mutagenesis of TMIE palmitoylation sites (C76C77A) |
Proceedings of the National Academy of Sciences of the United States of America |
High |
39999170
|
| 2009 |
In tmie zebrafish mutants (frameshift mutation), hair cells fail to incorporate FM1-43 and other fluorophores that traverse transduction channels, lack microphonic potentials, have short/disordered hair bundles, and stereocilia lack tip links and insertional plaques, establishing that TMIE is required for mechanotransduction channel function and tip-link integrity in hair cells. |
Positional cloning, FM1-43 dye uptake assay, microphonic potential recordings, electron/confocal microscopy of hair bundles in tmie zebrafish mutants |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19934034
|
| 2013 |
Hair cells of tmie-null (cir/cir) mice at postnatal day 3 (before hair-cell degeneration) fail to take up gentamicin, gentamicin-Texas Red conjugate, or FM1-43, compounds that enter hair cells through the mechanotransducer channel, demonstrating that TMIE is required for mechanotransducer channel activity. |
FM1-43 and gentamicin-Texas Red uptake assays in cir/cir vs control hair cells at P3 |
Comparative medicine |
Medium |
23582420
|
| 2002 |
Loss-of-function mutations in the mouse Tmie gene (40-kb deletion in spinner allele; nonsense mutation in a second allele) cause postnatal defects in cochlear hair cell stereocilia and profound failure to develop auditory function, establishing that Tmie is required for stereocilia integrity and mechanotransduction in mammalian hair cells. |
Positional cloning, genomic deletion characterization, auditory brainstem response, scanning/transmission electron microscopy of stereocilia in Tmie-null mice |
Human molecular genetics |
Medium |
12140191
|
| 2018 |
Ectopic expression of wild-type tmie transgene in cir/cir (Tmie-null) mice rescues hearing and vestibular behavior in a dose-dependent manner, with recovery correlating with cochlear transgene expression level, confirming that the tmie protein itself is the functional unit responsible for hearing. |
Transgenic rescue experiment in cir/cir mice; ABR and behavioral phenotype assessment correlated with cochlear transgene expression levels by Western blot |
Biochemical and biophysical research communications |
Medium |
18586001
|
| 2008 |
Tmie protein is expressed and localized to the stereocilia bundles of cochlear hair cells in the rat, with prominent expression in early postnatal stages, implicating it in stereocilia maturation; expression detected by immunostaining with a Tmie-specific antibody identifying a 17 kDa band on Western blot. |
Immunohistochemistry with anti-Tmie antibody, Western blot, RT-PCR in rat cochlea |
Histochemistry and cell biology |
Low |
18327602
|
| 2011 |
In a stable cell line expressing Myc-tagged Tmie, the protein localizes predominantly to the cellular membrane and to a lesser extent the cytoplasm, consistent with its predicted transmembrane domain architecture. |
Stable transfection of Myc-tagged Tmie, immunostaining with anti-Myc and anti-Tmie antibodies, Western blot |
Laboratory animal research |
Low |
22232643
|
| 2025 |
Slow adaptation of the mechanotransduction channel in mammalian cochlear and vestibular hair cells depends on PIP2 interactions with Tmie, independent of myosin motors. Slow adaptation is independent of myosin VIIa (upper tip-link motor), and exogenous PIP2 rescues slow adaptation when myosin motors are pharmacologically inhibited, supporting a model in which PIP2 binding by Tmie mediates slow adaptation at the lower end of the tip link. |
Electrophysiological recording of slow adaptation in cochlear and vestibular hair cells; pharmacological myosin inhibition; exogenous PIP2 application; analysis in Tmie mutant cells |
bioRxivpreprint |
Medium |
bio_10.1101_2025.04.01.646713
|