Structure of mouse protocadherin 15 of the stereocilia tip link in complex with LHFPL5

Ge, Jingpeng; Elferich, Johannes; Goehring, April; Zhao, Huaying; Schuck, Peter; Gouaux, Eric

doi:10.7554/elife.38770

Cited by 80 publications

(129 citation statements)

References 60 publications

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“…Another contribution comes from the ability of individual components to rebind after they rupture, dynamically maintaining the overall connection. Since CDH23 and PCDH15 both form strong cis-dimers in-vitro and in-vivo, facilitated by bonds along their ectodomains 6,7,[28][29][30][31] , the tip link may increase its lifetime through rebinding if opposing PCDH15 and CDH23 EC1-2 domains are kept in sufficiently close spatial proximity, creating a high effective local concentration and a fast on-rate.…”

Section: Main Textmentioning

confidence: 99%

“…Since our model predicts that the mechanical properties of tip-link cadherins significantly affect avidity, we made structural modifications to the tip link proteins to test this effect. Cis-dimerization links PCDH15 and CDH23 laterally at several points along their length 6,7,[28][29][30][31] , and may act to stabilize Ceff under force. We truncated PCDH15 and CDH23 to their first five EC domains in an effort to disrupt full cis-dimerization, and used single-molecule force spectroscopy to determine the effects on bond strength ( Fig.…”

Section: Main Textmentioning

confidence: 99%

“…9). Since CDH23 and PCDH15 are relatively stiff proteins 7,40,42 , are complexed with membrane and intracellular proteins 29,80,81 , and appear to be constrained at their transmembrane domains 11,12,82 , their N-termini may be constrained to a smaller volume and the effective concentration significantly higher, further increasing the re-forming rate. A more reasonable sphere of 50 nm gives an effective concentration of ~25 µM and a reforming time of just 0.29 s, 29 times faster than the unbinding rate at a resting tension of 10 pN.…”

Section: Turnover Of the Tip Link In Vivomentioning

confidence: 99%

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The Dynamic Strength of the Hair-Cell Tip Link Reveals Mechanisms of Hearing and Deafness

Mulhall

Ward

Yang

et al. 2019

Preprint

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Our senses of hearing and balance rely on the extraordinarily sensitive molecular machinery of the inner ear to convert deflections as small as the width of a single carbon atom 1,2 into electrical signals that the brain can process 3 . In humans and other vertebrates, transduction is mediated by hair cells 4 , where tension on tip links conveys force to mechanosensitive ion channels 5 . Each tip link comprises two helical filaments of atypical cadherins bound at their N-termini through two unique adhesion bonds 6-8 . Tip links must be strong enough to maintain a connection to the mechanotransduction channel under the dynamic forces exerted by sound or head movement-yet might also act as mechanical circuit breakers, releasing under extreme conditions to preserve the delicate structures within the hair cell. Previous studies have argued that this connection is exceptionally static, disrupted only by harsh chemical conditions or loud sound 9-12 . However, no direct mechanical measurements of the full tip-link connection have been performed. Here we describe the dynamics of the tiplink connection at single-molecule resolution and show how avidity conferred by its double stranded architecture enhances mechanical strength and lifetime, yet still enables it to act as a dynamic mechanical circuit breaker. We also show how the dynamic strength of the connection is facilitated by strong cisdimerization and tuned by extracellular Ca 2+ , and we describe the unexpected etiology of a hereditary human deafness mutation. Remarkably, the connection is several thousand times more dynamic than previously thought, challenging current assumptions about tip-link stability and turnover rate, and providing insight into how the mechanotransduction apparatus conveys mechanical information. Our results reveal fundamental mechanisms that underlie mechanoelectric transduction in the inner ear, and provide a foundation for studying multi-component linkages in other biological systems.

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Section: Main Textmentioning

confidence: 99%

Section: Main Textmentioning

confidence: 99%

Section: Turnover Of the Tip Link In Vivomentioning

confidence: 99%

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The Dynamic Strength of the Hair-Cell Tip Link Reveals Mechanisms of Hearing and Deafness

Mulhall

Ward

Yang

et al. 2019

Preprint

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“…On the contrary, a longer and straight δ1 complex might prevent adhesion mediated by shorter and tilted classical complexes. Determining how protocadherins orient with respect to the membrane plane, as recently explored for protocadherin-15 57 , will be critical to elucidate their function.…”

Section: Discussionmentioning

confidence: 99%

An Adhesive Interface for the Non-Clustered δ1 Protocadherin-1 Involved in Respiratory Diseases

Modak

Sotomayor

2018

Preprint

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150 WORDS) 1 Cadherins form a large family of calcium-dependent adhesive proteins involved in 2 morphogenesis, cell differentiation, and neuronal connectivity. Non-clustered δ1 protocadherins 3 form a cadherin subgroup of proteins with seven extracellular cadherin (EC) repeats and 4 cytoplasmic domains distinct from those of classical cadherins. The non-clustered δ1 5 protocadherins mediate homophilic adhesion and have been implicated in various diseases 6including asthma, autism, and cancer. Here we present X-ray crystal structures of Protocadherin-7 1 (PCDH1), a δ1-protocadherin member essential for New World hantavirus infection that is 8 typically expressed in the brain, airway epithelium, skin keratinocytes, and lungs. The structures 9 suggest a binding mode that involves antiparallel overlap of repeats EC1 to EC4. Mutagenesis 10 combined with binding assays and biochemical experiments validated this mode of adhesion. 11 Overall, these results reveal the molecular mechanism underlying adhesiveness of PCDH1 and 12 δ1-protocadherins, also shedding light on PCDH1's role in maintaining airway epithelial 13 integrity, the loss of which causes respiratory diseases. 14 15

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“…Our current understanding is that the proteins of the MET complex consist of the tip link 41 proteins cadherin 23 (CDH23) and protocadherin 15 (PCDH15) at the upper and lower ends of the tip 42 link respectively (Kazmierczak et al, 2007), the pore-forming subunits transmembrane channel-like 43 proteins TMC1 and TMC2 (Kawashima et al, 2011;Pan et al, 2013Pan et al, , 2018, and the accessory subunits 44 transmembrane inner ear (TMIE) and lipoma HMGIC fusion partner-like 5 (LHFPL5) (Cunningham 45 and immunoprecipitation experiments in heterologous cells suggests that LHFPL5 can directly interact 61 with PCDH15 and TMIE (Xiong et al, 2012;Zhao et al, 2014). A structure of the PCDH15 -LHFPL5 62 complex has also been reported (Ge et al, 2018). 63 A precise role for LHFPL5 has yet to be defined.…”

Section: Introduction 38mentioning

confidence: 99%

The lhfpl5 ohnologs lhfpl5a and lhfpl5b are required for mechanotransduction in distinct populations of sensory hair cells in zebrafish

Erickson

Pacentine²,

Venuto

et al. 2019

Preprint

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Article type 15Original Research 16 1Abstract 17 Hair cells sense and transmit auditory, vestibular, and hydrodynamic information by converting 18 mechanical stimuli into electrical signals. This process of mechano-electrical transduction (MET) 19 requires a mechanically-gated channel localized in the apical stereocilia of hair cells. In mice, lipoma 20 HMGIC fusion partner-like 5 (LHFPL5) acts as an auxiliary subunit of the MET channel whose 21 primary role is to correctly localize PCDH15 and TMC1 to the mechanotransduction complex. 22Zebrafish have two lhfpl5 genes (lhfpl5a and lhfpl5b), but their individual contributions to MET 23 channel assembly and function have not been analyzed. 24Here we show that the zebrafish lhfpl5 genes are expressed in discrete populations of hair cells: lhfpl5a 25 expression is restricted to auditory and vestibular hair cells in the inner ear, while lhfpl5b expression 26 is specific to hair cells of the lateral line organ. Consequently, lhfpl5a mutants exhibit defects in 27 auditory and vestibular function, while disruption of lhfpl5b affects hair cells only in the lateral line 28 neuromasts. In contrast to previous reports in mice, localization of Tmc1 does not depend upon Lhfpl5 29 function in either the inner ear or lateral line organ. In both lhfpl5a and lhfpl5b mutants, GFP-tagged 30Tmc1 and Tmc2b proteins still localize to the stereocilia of hair cells. Using a stably integrated GFP-31Lhfpl5a transgene, we show that the tip link cadherins Pcdh15a and Cdh23, along with the Myo7aa 32 motor protein, are required for correct Lhfpl5a localization at the tips of stereocilia. Our work 33corroborates the evolutionarily conserved co-dependence between Lhfpl5 and Pcdh15, but also reveals 34 novel requirements for Cdh23 and Myo7aa to correctly localize Lhfpl5a. In addition, our data suggest 35 that targeting of Tmc1 and Tmc2b proteins to stereocilia in zebrafish hair cells occurs independently 36 of Lhfpl5 proteins. 37

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Structure of mouse protocadherin 15 of the stereocilia tip link in complex with LHFPL5

Cited by 80 publications

References 60 publications

The Dynamic Strength of the Hair-Cell Tip Link Reveals Mechanisms of Hearing and Deafness

The Dynamic Strength of the Hair-Cell Tip Link Reveals Mechanisms of Hearing and Deafness

An Adhesive Interface for the Non-Clustered δ1 Protocadherin-1 Involved in Respiratory Diseases

The lhfpl5 ohnologs lhfpl5a and lhfpl5b are required for mechanotransduction in distinct populations of sensory hair cells in zebrafish

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