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Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding
Mutations in myosin VI have been associated with autosomal-recessive (DFNB37) and autosomal-dominant (DFNA22) deafness in humans. Here, we characterise an myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to the deletio...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5074368/ https://www.ncbi.nlm.nih.gov/pubmed/27474411 http://dx.doi.org/10.1042/BCJ20160571 |
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author | Arden, Susan D. Tumbarello, David A. Butt, Tariq Kendrick-Jones, John Buss, Folma |
author_facet | Arden, Susan D. Tumbarello, David A. Butt, Tariq Kendrick-Jones, John Buss, Folma |
author_sort | Arden, Susan D. |
collection | PubMed |
description | Mutations in myosin VI have been associated with autosomal-recessive (DFNB37) and autosomal-dominant (DFNA22) deafness in humans. Here, we characterise an myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to the 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. |
format | Online Article Text |
id | pubmed-5074368 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-50743682016-10-21 Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding Arden, Susan D. Tumbarello, David A. Butt, Tariq Kendrick-Jones, John Buss, Folma Biochem J Research Articles Mutations in myosin VI have been associated with autosomal-recessive (DFNB37) and autosomal-dominant (DFNA22) deafness in humans. Here, we characterise an myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to the 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. Portland Press Ltd. 2016-10-01 2016-09-27 /pmc/articles/PMC5074368/ /pubmed/27474411 http://dx.doi.org/10.1042/BCJ20160571 Text en © 2016 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0) . |
spellingShingle | Research Articles Arden, Susan D. Tumbarello, David A. Butt, Tariq Kendrick-Jones, John Buss, Folma Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title | Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title_full | Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title_fullStr | Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title_full_unstemmed | Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title_short | Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding |
title_sort | loss of cargo binding in the human myosin vi deafness mutant (r1166x) leads to increased actin filament binding |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5074368/ https://www.ncbi.nlm.nih.gov/pubmed/27474411 http://dx.doi.org/10.1042/BCJ20160571 |
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