The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions

The formation and maintenance of the mitotic spindle during cell division requires several microtubule-interacting motor proteins. Members of the kinesin-5 family play an essential role in the bipolar organization of the spindle. These highly conserved, homotetrameric proteins cross-link anti-parall...

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Autores principales:	Tobias Strauß, Saskia Schattner, Stefan Hoth, Wilhelm J. Walter
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	kinesin <i>Arabidopsis thaliana</i> motor protein intracellular transport single-molecule Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/718f54dd836c4c81a6c6294113b6380a
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spelling	oai:doaj.org-article:718f54dd836c4c81a6c6294113b6380a2021-11-11T16:50:44ZThe <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions10.3390/ijms2221113611422-00671661-6596https://doaj.org/article/718f54dd836c4c81a6c6294113b6380a2021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11361https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067The formation and maintenance of the mitotic spindle during cell division requires several microtubule-interacting motor proteins. Members of the kinesin-5 family play an essential role in the bipolar organization of the spindle. These highly conserved, homotetrameric proteins cross-link anti-parallel microtubules and slide them apart to elongate the spindle during the equal separation of chromosomes. Whereas vertebrate kinesin-5 proteins are well studied, knowledge about the biochemical properties and the function of plant kinesin-5 proteins is still limited. Here, we characterized the properties of AtKRP125b, one of four kinesin-5 proteins in <i>Arabidopsis thaliana</i>. In in vitro motility assays, AtKRP125b displayed the archetypal characteristics of a kinesin-5 protein, a low velocity of about 20 nm·s<sup>−1</sup>, and a plus end-directed, processive movement. Moreover, AtKRP125b was able to cross-link microtubules and to slide them apart, as required for developing and maintaining the mitotic spindle. In line with such a function, GFP-AtKRP125b fusion proteins were predominantly detected in the nucleus when expressed in <i>Arabidopsis thaliana</i> leaf protoplasts or <i>Nicotiana benthamiana</i> epidermis cells and analyzed by confocal microscopy. However, we also detected GFP signals in the cytoplasm, suggesting additional functions. By generating and analyzing <i>AtKRP125b</i> promoter-reporter lines, we showed that the <i>AtKRP125b</i> promoter was active in the vascular tissue of roots, lateral roots, cotyledons, and true leaves. Remarkably, we could not detect promoter activity in meristematic tissues. Taken together, our biochemical data support a role of AtKRP125b in mitosis, but it may also have additional functions outside the nucleus and during interphase.Tobias StraußSaskia SchattnerStefan HothWilhelm J. WalterMDPI AGarticlekinesin<i>Arabidopsis thaliana</i>motor proteinintracellular transportsingle-moleculeBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11361, p 11361 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	kinesin <i>Arabidopsis thaliana</i> motor protein intracellular transport single-molecule Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	kinesin <i>Arabidopsis thaliana</i> motor protein intracellular transport single-molecule Biology (General) QH301-705.5 Chemistry QD1-999 Tobias Strauß Saskia Schattner Stefan Hoth Wilhelm J. Walter The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
description	The formation and maintenance of the mitotic spindle during cell division requires several microtubule-interacting motor proteins. Members of the kinesin-5 family play an essential role in the bipolar organization of the spindle. These highly conserved, homotetrameric proteins cross-link anti-parallel microtubules and slide them apart to elongate the spindle during the equal separation of chromosomes. Whereas vertebrate kinesin-5 proteins are well studied, knowledge about the biochemical properties and the function of plant kinesin-5 proteins is still limited. Here, we characterized the properties of AtKRP125b, one of four kinesin-5 proteins in <i>Arabidopsis thaliana</i>. In in vitro motility assays, AtKRP125b displayed the archetypal characteristics of a kinesin-5 protein, a low velocity of about 20 nm·s<sup>−1</sup>, and a plus end-directed, processive movement. Moreover, AtKRP125b was able to cross-link microtubules and to slide them apart, as required for developing and maintaining the mitotic spindle. In line with such a function, GFP-AtKRP125b fusion proteins were predominantly detected in the nucleus when expressed in <i>Arabidopsis thaliana</i> leaf protoplasts or <i>Nicotiana benthamiana</i> epidermis cells and analyzed by confocal microscopy. However, we also detected GFP signals in the cytoplasm, suggesting additional functions. By generating and analyzing <i>AtKRP125b</i> promoter-reporter lines, we showed that the <i>AtKRP125b</i> promoter was active in the vascular tissue of roots, lateral roots, cotyledons, and true leaves. Remarkably, we could not detect promoter activity in meristematic tissues. Taken together, our biochemical data support a role of AtKRP125b in mitosis, but it may also have additional functions outside the nucleus and during interphase.
format	article
author	Tobias Strauß Saskia Schattner Stefan Hoth Wilhelm J. Walter
author_facet	Tobias Strauß Saskia Schattner Stefan Hoth Wilhelm J. Walter
author_sort	Tobias Strauß
title	The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
title_short	The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
title_full	The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
title_fullStr	The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
title_full_unstemmed	The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions
title_sort	<i>arabidopsis thaliana</i> kinesin-5 atkrp125b is a processive, microtubule-sliding motor protein with putative plant-specific functions
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/718f54dd836c4c81a6c6294113b6380a
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The <i>Arabidopsis thaliana</i> Kinesin-5 AtKRP125b Is a Processive, Microtubule-Sliding Motor Protein with Putative Plant-Specific Functions

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