Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://doi.org/10.1242/jcs.186064 |
Resumo: | Microtubules and F-actin, and their associated motor proteins, are considered to play complementary roles in long- and short-range organelle transport. However, there is growing appreciation that myosin/F-actin networks can drive long-range transport. In melanocytes, myosin-Va and kinesin-1 have both been proposed as long-range centrifugal transporters moving melanosomes into the peripheral dendrites. Here,we investigated the role of kinesin-1 heavy chain (Kif5b) and its suggested targeting factor Rab1a in transport. We performed confocal microscopy and subcellular fractionation, but did not detect Kif5b or Rab1a on melanosomes. Meanwhile functional studies, using siRNA knockdown and dominant negative mutants, did not support a role for Kif5b or Rab1a in melanosome transport. To probe the potential of Kif5b to function in transport, we generated fusion proteins that target active Kif5b to melanosomes and tested their ability to rescue perinuclear clustering in myosin-Va-deficient cells. Expression of these chimeras, but not full-length Kif5b, dispersed melanosomes with similar efficiency to myosin-Va. Our data indicate that kinesin and microtubules can compensate for defects in myosin-Va and actin-based transport in mammals, but that endogenous Kif5b does not have an important role in transport of melanocytes due to its inefficient recruitment to melanosomes. © 2017. Published by The Company of Biologists Ltd. |
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Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transportActinKinesin-1MelanocyteMicrotubulesMyosin-VaOrganelle transportMicrotubules and F-actin, and their associated motor proteins, are considered to play complementary roles in long- and short-range organelle transport. However, there is growing appreciation that myosin/F-actin networks can drive long-range transport. In melanocytes, myosin-Va and kinesin-1 have both been proposed as long-range centrifugal transporters moving melanosomes into the peripheral dendrites. Here,we investigated the role of kinesin-1 heavy chain (Kif5b) and its suggested targeting factor Rab1a in transport. We performed confocal microscopy and subcellular fractionation, but did not detect Kif5b or Rab1a on melanosomes. Meanwhile functional studies, using siRNA knockdown and dominant negative mutants, did not support a role for Kif5b or Rab1a in melanosome transport. To probe the potential of Kif5b to function in transport, we generated fusion proteins that target active Kif5b to melanosomes and tested their ability to rescue perinuclear clustering in myosin-Va-deficient cells. Expression of these chimeras, but not full-length Kif5b, dispersed melanosomes with similar efficiency to myosin-Va. Our data indicate that kinesin and microtubules can compensate for defects in myosin-Va and actin-based transport in mammals, but that endogenous Kif5b does not have an important role in transport of melanocytes due to its inefficient recruitment to melanosomes. © 2017. Published by The Company of Biologists Ltd.Centro de Estudos de Doenças Crónicas (CEDOC)NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)RUNRobinson, C.L.Evans, R.D.Briggs, D.A.Ramalho, J.S.Hume, A.N.2017-10-25T22:00:16Z2017-06-152017-06-15T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10application/pdfhttps://doi.org/10.1242/jcs.186064eng0021-9533PURE: 3228880https://www.scopus.com/inward/record.uri?eid=2-s2.0-85020509919&doi=10.1242%2fjcs.186064&partnerID=40&md5=8376884ac605d19afee9c3af4a700b5chttps://doi.org/10.1242/jcs.186064info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:12:46Zoai:run.unl.pt:10362/24606Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:28:04.799949Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
title |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
spellingShingle |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport Robinson, C.L. Actin Kinesin-1 Melanocyte Microtubules Myosin-Va Organelle transport |
title_short |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
title_full |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
title_fullStr |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
title_full_unstemmed |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
title_sort |
Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport |
author |
Robinson, C.L. |
author_facet |
Robinson, C.L. Evans, R.D. Briggs, D.A. Ramalho, J.S. Hume, A.N. |
author_role |
author |
author2 |
Evans, R.D. Briggs, D.A. Ramalho, J.S. Hume, A.N. |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Centro de Estudos de Doenças Crónicas (CEDOC) NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) RUN |
dc.contributor.author.fl_str_mv |
Robinson, C.L. Evans, R.D. Briggs, D.A. Ramalho, J.S. Hume, A.N. |
dc.subject.por.fl_str_mv |
Actin Kinesin-1 Melanocyte Microtubules Myosin-Va Organelle transport |
topic |
Actin Kinesin-1 Melanocyte Microtubules Myosin-Va Organelle transport |
description |
Microtubules and F-actin, and their associated motor proteins, are considered to play complementary roles in long- and short-range organelle transport. However, there is growing appreciation that myosin/F-actin networks can drive long-range transport. In melanocytes, myosin-Va and kinesin-1 have both been proposed as long-range centrifugal transporters moving melanosomes into the peripheral dendrites. Here,we investigated the role of kinesin-1 heavy chain (Kif5b) and its suggested targeting factor Rab1a in transport. We performed confocal microscopy and subcellular fractionation, but did not detect Kif5b or Rab1a on melanosomes. Meanwhile functional studies, using siRNA knockdown and dominant negative mutants, did not support a role for Kif5b or Rab1a in melanosome transport. To probe the potential of Kif5b to function in transport, we generated fusion proteins that target active Kif5b to melanosomes and tested their ability to rescue perinuclear clustering in myosin-Va-deficient cells. Expression of these chimeras, but not full-length Kif5b, dispersed melanosomes with similar efficiency to myosin-Va. Our data indicate that kinesin and microtubules can compensate for defects in myosin-Va and actin-based transport in mammals, but that endogenous Kif5b does not have an important role in transport of melanocytes due to its inefficient recruitment to melanosomes. © 2017. Published by The Company of Biologists Ltd. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-10-25T22:00:16Z 2017-06-15 2017-06-15T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://doi.org/10.1242/jcs.186064 |
url |
https://doi.org/10.1242/jcs.186064 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
0021-9533 PURE: 3228880 https://www.scopus.com/inward/record.uri?eid=2-s2.0-85020509919&doi=10.1242%2fjcs.186064&partnerID=40&md5=8376884ac605d19afee9c3af4a700b5c https://doi.org/10.1242/jcs.186064 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
10 application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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