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Loss of cargo binding in the human myosin VI deafness mutant (R1166X) increases actin filament binding

Loss of cargo binding in the human myosin VI deafness mutant (R1166X) increases actin filament binding
Loss of cargo binding in the human myosin VI deafness mutant (R1166X) increases actin filament binding
Mutations in myosin VI have been associated with autosomal recessive (DFNB37) and autosomal dominant (DFNA22) deafness in humans. Here, we characterise a myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to deletion of the C-terminal 120 amino acids of the myosin VI cargo-binding domain, which includes the WWY binding motif for the adaptor proteins LMTK2, Tom1 as well as Dab2. Interestingly, compromising myosin VI vesicle binding ability by expressing myosin VI with the R1166X mutation or with single point mutations in the adaptor binding sites leads to increased F-actin-binding of this myosin in vitro and in vivo As our results highlight the importance of cargo attachment for regulating actin binding to the motor domain, we perform a detailed characterisation of adaptor protein binding and identify single amino acids within myosin VI required for binding to cargo adaptors. We not only show that the adaptor proteins can directly interact with the cargo-binding tail of myosin VI, but our in vitro studies also suggest that multiple adaptor proteins can bind simultaneously to non-overlapping sites in the myosin VI tail. In conclusion, our characterisation of the human myosin VI deafness mutant (R1166X) suggests that defects in cargo binding may leave myosin VI in a primed/activated state with an increased actin-binding ability
1470-8728
1-20
Arden, Susan D.
47286333-3c42-4c14-ac71-14c5b7ac4e61
Tumbarello, David A.
75c6932e-fdbf-4d3c-bb4f-48fbbdba93a2
Butt, Tariq
7135d4f6-93fa-45b2-abab-fb93a7fb127c
Kendrick-Jones, John
0c756304-5044-44a0-8277-0e8b352b8aae
Buss, Folma
f8b3fde3-6724-4412-aa65-674cbfb73121
Arden, Susan D.
47286333-3c42-4c14-ac71-14c5b7ac4e61
Tumbarello, David A.
75c6932e-fdbf-4d3c-bb4f-48fbbdba93a2
Butt, Tariq
7135d4f6-93fa-45b2-abab-fb93a7fb127c
Kendrick-Jones, John
0c756304-5044-44a0-8277-0e8b352b8aae
Buss, Folma
f8b3fde3-6724-4412-aa65-674cbfb73121

Arden, Susan D., Tumbarello, David A., Butt, Tariq, Kendrick-Jones, John and Buss, Folma (2016) Loss of cargo binding in the human myosin VI deafness mutant (R1166X) increases actin filament binding. Biochemical Journal, 1-20. (doi:10.1042/BCJ20160571). (PMID:27474411)

Record type: Article

Abstract

Mutations in myosin VI have been associated with autosomal recessive (DFNB37) and autosomal dominant (DFNA22) deafness in humans. Here, we characterise a myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to deletion of the C-terminal 120 amino acids of the myosin VI cargo-binding domain, which includes the WWY binding motif for the adaptor proteins LMTK2, Tom1 as well as Dab2. Interestingly, compromising myosin VI vesicle binding ability by expressing myosin VI with the R1166X mutation or with single point mutations in the adaptor binding sites leads to increased F-actin-binding of this myosin in vitro and in vivo As our results highlight the importance of cargo attachment for regulating actin binding to the motor domain, we perform a detailed characterisation of adaptor protein binding and identify single amino acids within myosin VI required for binding to cargo adaptors. We not only show that the adaptor proteins can directly interact with the cargo-binding tail of myosin VI, but our in vitro studies also suggest that multiple adaptor proteins can bind simultaneously to non-overlapping sites in the myosin VI tail. In conclusion, our characterisation of the human myosin VI deafness mutant (R1166X) suggests that defects in cargo binding may leave myosin VI in a primed/activated state with an increased actin-binding ability

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Accepted/In Press date: 28 July 2016
e-pub ahead of print date: 29 July 2016
Published date: 27 September 2016
Organisations: Centre for Biological Sciences

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Local EPrints ID: 398704
URI: https://eprints.soton.ac.uk/id/eprint/398704
ISSN: 1470-8728
PURE UUID: 3334c2f5-e560-451f-baa8-86b9472a8fc1
ORCID for David A. Tumbarello: ORCID iD orcid.org/0000-0002-5169-0561

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Date deposited: 08 Aug 2016 08:01
Last modified: 06 Jun 2018 12:23

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Contributors

Author: Susan D. Arden
Author: Tariq Butt
Author: John Kendrick-Jones
Author: Folma Buss

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