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A ‘molecular guillotine’ reveals the interphase function of Kinesin-5
Motor proteins are important for transport and force generation in a variety of cellular processes and in morphogenesis. Here, we describe a general strategy for conditional motor mutants by inserting a protease cleavage site into the ‘neck’ between the head domain and the stalk of the motor protein...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5826049/ https://www.ncbi.nlm.nih.gov/pubmed/29361546 http://dx.doi.org/10.1242/jcs.210583 |
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author | Lv, Zhiyi Rosenbaum, Jan Aspelmeier, Timo Großhans, Jörg |
author_facet | Lv, Zhiyi Rosenbaum, Jan Aspelmeier, Timo Großhans, Jörg |
author_sort | Lv, Zhiyi |
collection | PubMed |
description | Motor proteins are important for transport and force generation in a variety of cellular processes and in morphogenesis. Here, we describe a general strategy for conditional motor mutants by inserting a protease cleavage site into the ‘neck’ between the head domain and the stalk of the motor protein, making the protein susceptible to proteolytic cleavage at the neck by the corresponding protease. To demonstrate the feasibility of this approach, we inserted the cleavage site of the tobacco etch virus (TEV) protease into the neck of the tetrameric motor Kinesin-5. Application of TEV protease led to a specific depletion and functional loss of Kinesin-5 in Drosophila embryos. With our approach, we revealed that Kinesin-5 stabilizes the microtubule network during interphase in syncytial embryos. The ‘molecular guillotine’ can potentially be applied to many motor proteins because Kinesins and myosins have conserved structures with accessible neck regions. This article has an associated First Person interview with the first author of the paper. |
format | Online Article Text |
id | pubmed-5826049 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-58260492018-04-11 A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 Lv, Zhiyi Rosenbaum, Jan Aspelmeier, Timo Großhans, Jörg J Cell Sci Tools and Resources Motor proteins are important for transport and force generation in a variety of cellular processes and in morphogenesis. Here, we describe a general strategy for conditional motor mutants by inserting a protease cleavage site into the ‘neck’ between the head domain and the stalk of the motor protein, making the protein susceptible to proteolytic cleavage at the neck by the corresponding protease. To demonstrate the feasibility of this approach, we inserted the cleavage site of the tobacco etch virus (TEV) protease into the neck of the tetrameric motor Kinesin-5. Application of TEV protease led to a specific depletion and functional loss of Kinesin-5 in Drosophila embryos. With our approach, we revealed that Kinesin-5 stabilizes the microtubule network during interphase in syncytial embryos. The ‘molecular guillotine’ can potentially be applied to many motor proteins because Kinesins and myosins have conserved structures with accessible neck regions. This article has an associated First Person interview with the first author of the paper. The Company of Biologists Ltd 2018-02-01 /pmc/articles/PMC5826049/ /pubmed/29361546 http://dx.doi.org/10.1242/jcs.210583 Text en © 2018. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Tools and Resources Lv, Zhiyi Rosenbaum, Jan Aspelmeier, Timo Großhans, Jörg A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title | A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title_full | A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title_fullStr | A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title_full_unstemmed | A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title_short | A ‘molecular guillotine’ reveals the interphase function of Kinesin-5 |
title_sort | ‘molecular guillotine’ reveals the interphase function of kinesin-5 |
topic | Tools and Resources |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5826049/ https://www.ncbi.nlm.nih.gov/pubmed/29361546 http://dx.doi.org/10.1242/jcs.210583 |
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