Inefficient recruitment of kinesin-1 to melanosomes precludes it from facilitating their transport

Detalhes bibliográficos
Autor(a) principal: Robinson, C.L.
Data de Publicação: 2017
Outros Autores: Evans, R.D., Briggs, D.A., Ramalho, J.S., Hume, A.N.
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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spelling 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
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https://doi.org/10.1242/jcs.186064
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