Cargando…

Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity

The assembly and functionality of the mitotic spindle depends on the coordinated activities of microtubule-associated motor proteins of the dynein and kinesin superfamily. Our current understanding of the function of motor proteins is significantly shaped by studies using Xenopus laevis egg extract...

Descripción completa

Detalles Bibliográficos
Autores principales: Möckel, Martin M., Heim, Andreas, Tischer, Thomas, Mayer, Thomas U.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5399559/
https://www.ncbi.nlm.nih.gov/pubmed/28228376
http://dx.doi.org/10.1242/bio.023952
_version_ 1783230662670548992
author Möckel, Martin M.
Heim, Andreas
Tischer, Thomas
Mayer, Thomas U.
author_facet Möckel, Martin M.
Heim, Andreas
Tischer, Thomas
Mayer, Thomas U.
author_sort Möckel, Martin M.
collection PubMed
description The assembly and functionality of the mitotic spindle depends on the coordinated activities of microtubule-associated motor proteins of the dynein and kinesin superfamily. Our current understanding of the function of motor proteins is significantly shaped by studies using Xenopus laevis egg extract as its open structure allows complex experimental manipulations hardly feasible in other model systems. Yet, the Kinesin-8 orthologue of human Kif18A has not been described in Xenopus laevis so far. Here, we report the cloning and characterization of Xenopus laevis (Xl) Kif18A. Xenopus Kif18A is expressed during oocyte maturation and its depletion from meiotic egg extract results in severe spindle defects. These defects can be rescued by wild-type Kif18A, but not Kif18A lacking motor activity or the C-terminus. Single-molecule microscopy assays revealed that Xl_Kif18A possesses high processivity, which depends on an additional C-terminal microtubule-binding site. Human tissue culture cells depleted of endogenous Kif18A display mitotic defects, which can be rescued by wild-type, but not tail-less Xl_Kif18A. Thus, Xl_Kif18A is the functional orthologue of human Kif18A whose activity is essential for the correct function of meiotic spindles in Xenopus oocytes.
format Online
Article
Text
id pubmed-5399559
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher The Company of Biologists Ltd
record_format MEDLINE/PubMed
spelling pubmed-53995592017-05-02 Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity Möckel, Martin M. Heim, Andreas Tischer, Thomas Mayer, Thomas U. Biol Open Research Article The assembly and functionality of the mitotic spindle depends on the coordinated activities of microtubule-associated motor proteins of the dynein and kinesin superfamily. Our current understanding of the function of motor proteins is significantly shaped by studies using Xenopus laevis egg extract as its open structure allows complex experimental manipulations hardly feasible in other model systems. Yet, the Kinesin-8 orthologue of human Kif18A has not been described in Xenopus laevis so far. Here, we report the cloning and characterization of Xenopus laevis (Xl) Kif18A. Xenopus Kif18A is expressed during oocyte maturation and its depletion from meiotic egg extract results in severe spindle defects. These defects can be rescued by wild-type Kif18A, but not Kif18A lacking motor activity or the C-terminus. Single-molecule microscopy assays revealed that Xl_Kif18A possesses high processivity, which depends on an additional C-terminal microtubule-binding site. Human tissue culture cells depleted of endogenous Kif18A display mitotic defects, which can be rescued by wild-type, but not tail-less Xl_Kif18A. Thus, Xl_Kif18A is the functional orthologue of human Kif18A whose activity is essential for the correct function of meiotic spindles in Xenopus oocytes. The Company of Biologists Ltd 2017-02-22 /pmc/articles/PMC5399559/ /pubmed/28228376 http://dx.doi.org/10.1242/bio.023952 Text en © 2017. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0This 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 Research Article
Möckel, Martin M.
Heim, Andreas
Tischer, Thomas
Mayer, Thomas U.
Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title_full Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title_fullStr Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title_full_unstemmed Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title_short Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity
title_sort xenopus laevis kif18a is a highly processive kinesin required for meiotic spindle integrity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5399559/
https://www.ncbi.nlm.nih.gov/pubmed/28228376
http://dx.doi.org/10.1242/bio.023952
work_keys_str_mv AT mockelmartinm xenopuslaeviskif18aisahighlyprocessivekinesinrequiredformeioticspindleintegrity
AT heimandreas xenopuslaeviskif18aisahighlyprocessivekinesinrequiredformeioticspindleintegrity
AT tischerthomas xenopuslaeviskif18aisahighlyprocessivekinesinrequiredformeioticspindleintegrity
AT mayerthomasu xenopuslaeviskif18aisahighlyprocessivekinesinrequiredformeioticspindleintegrity