The TMC1 and TMC2 membrane proteins are critical components of the transduction complex in vertebrate hair cells, which converts the vibration of sound to an electrical signal, and it is likely they are central subunits of the long-sought transduction channel. However little is known about these proteins at a molecular level, other than their primary amino acid sequence. In this project we will determine the structure of TMC channels at atomic resolution to determine their topology, stoichiometry, permeation pathway, gating, and interaction with other subunits of the complex. We will first use biochemical methods to determine both the stoichiometry of the native channel, and the interaction of TMC1 and TMC2 with each other and with the other six members of the TMC family. We will then synthesize and purify a TMC channel, and determine its structure with electron microscopy. Low-resolution EM structures will confirm stoichiometry and tertiary structure, and may reveal a pore domain. High-resolution cryo-EM images will enable the solution of atomic structures of TMCs in different states. This will reveal a putative pore domain and possible gating movements. Finally, we will test the structure with cysteine mutagenesis of TMC1 and physiological recording. Individual amino acids will be replaced with cysteine and the cys-mutant TMC1 will be expressed in hair cells lacking wild-type TMC1 and TMC2. Cysteine-modifying reagents will be applied during physiological recording; their effects on permeation and gating will be correlated with the predicted structure. Solution of the TMC structure will answer questions about the mechanism of hearing that have stood for over 50 years. It will accelerate our understanding of how the entire transduction complex assembles and functions, and it will lead to molecular understanding of inherited deafness.
The molecular core of human hearing is the transduction channel complex that is opened by the mechanical stimulus of sound and that enables electrical current flow to initiate a neural signal. This complex is composed of several proteins including TMC1 and TMC2. Understanding their atomic structure will solve one of the fundamental questions in auditory science, and will both reveal how inherited mutations in these proteins cause deafness and suggest how such deafness might be treated.
|Pan, Bifeng; Akyuz, Nurunisa; Liu, Xiao-Ping et al. (2018) TMC1 Forms the Pore of Mechanosensory Transduction Channels in Vertebrate Inner Ear Hair Cells. Neuron 99:736-753.e6|
|Corey, David P; Akyuz, Nurunisa; Holt, Jeffrey R (2018) Function and Dysfunction of TMC Channels in Inner Ear Hair Cells. Cold Spring Harb Perspect Med :|
|Pan, Bifeng; Askew, Charles; Galvin, Alice et al. (2017) Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c. Nat Biotechnol 35:264-272|
|Delling, M; Indzhykulian, A A; Liu, X et al. (2016) Primary cilia are not calcium-responsive mechanosensors. Nature 531:656-60|
|Asadnia, Mohsen; Kottapalli, Ajay Giri Prakash; Karavitaki, K Domenica et al. (2016) From Biological Cilia to Artificial Flow Sensors: Biomimetic Soft Polymer Nanosensors with High Sensing Performance. Sci Rep 6:32955|
|Corey, David P; Holt, Jeffrey R (2016) Are TMCs the Mechanotransduction Channels of Vertebrate Hair Cells? J Neurosci 36:10921-10926|
|Vogl, Christian; Panou, Iliana; Yamanbaeva, Gulnara et al. (2016) Tryptophan-rich basic protein (WRB) mediates insertion of the tail-anchored protein otoferlin and is required for hair cell exocytosis and hearing. EMBO J 35:2536-2552|
|Wu, Xudong; Indzhykulian, Artur A; Niksch, Paul D et al. (2016) Hair-Cell Mechanotransduction Persists in TRP Channel Knockout Mice. PLoS One 11:e0155577|
|Christensen, Adam P; Akyuz, Nurunisa; Corey, David P (2016) The Outer Pore and Selectivity Filter of TRPA1. PLoS One 11:e0166167|
|Kwan, Kelvin Y; Shen, Jun; Corey, David P (2015) C-MYC transcriptionally amplifies SOX2 target genes to regulate self-renewal in multipotent otic progenitor cells. Stem Cell Reports 4:47-60|
Showing the most recent 10 out of 28 publications