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Open AccessJournal ArticleDOI

The Chromokinesin Kid Is Required for Maintenance of Proper Metaphase Spindle Size

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TLDR
It is suggested that distinct from its role in chromosome movement, Kid contributes to spindle morphogenesis by mediating spindle microtubules stabilization.
Abstract
The human chromokinesin Kid/kinesin-10, a plus end-directed microtubule (MT)-based motor with both microtubule- and DNA-binding domains, is required for proper chromosome alignment at the metaphase plate. Here, we performed RNA interference experiments to deplete endogenous Kid from HeLa cells and confirmed defects in metaphase chromosome arm alignment in Kid-depleted cells. In addition, we noted a shortening of the spindle length, resulting in a pole-to-pole distance only 80% of wild type. The spindle microtubule-bundles with which Kid normally colocalize became less robust. Rescue of the two Kid deficiency phenotypes-imprecise chromosome alignment at metaphase and shortened spindles- exhibited distinct requirements. Mutants lacking either the DNA-binding domain or the MT motor ATPase failed to rescue the former defect, whereas rescue of the shortened spindle phenotype required neither activity. Kid also exhibits microtubule bundling activity in vitro, and rescue of the shortened spindle phenotype and the bundling activity displayed similar domain requirements, except that rescue required a coiled-coil domain not needed for bundling. These results suggest that distinct from its role in chromosome movement, Kid contributes to spindle morphogenesis by mediating spindle microtubules stabilization.

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Kinesins and cancer

TL;DR: Three compounds that inhibit two mitotic kinesins (EG5 (also known as KIF11) and centromere-associated protein E (CENPE) have entered clinical trials either as monotherapies or in combination with other drugs.
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Profiling the human protein-DNA interactome reveals ERK2 as a transcriptional repressor of interferon signaling.

TL;DR: A combined bioinformatics and protein microarray-based strategy is used to systematically characterize the human protein-DNA interactome, identifying 17,718 PDIs between 460 DNA motifs predicted to regulate transcription and 4,191 human proteins of various functional classes.
Book ChapterDOI

Mechanisms of mitotic spindle assembly and function.

TL;DR: A detailed account of the key observations leading to current models of mitotic spindle assembly can be found in this article, as well as an up-to-date status report on this exciting field.
Journal ArticleDOI

The spatial arrangement of chromosomes during prometaphase facilitates spindle assembly

TL;DR: Spindle formation involves a previously overlooked stage of chromosome prepositioning which promotes formation of amphitelic attachments, and a computational model predicts that this toroidal distribution of chromosomes exposes kinetochores to a high density of microtubules which facilitates subsequent formation of amphibian attachments.
Journal ArticleDOI

Force and Length in the Mitotic Spindle

TL;DR: Current understanding of the basic architecture and dynamics of the metaphase spindle, and some of the elementary force-producing mechanisms, are reviewed, and models for force integration and spindle length determination are discussed.
References
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Journal ArticleDOI

Small Molecule Inhibitor of Mitotic Spindle Bipolarity Identified in a Phenotype-Based Screen

TL;DR: In vitro, monastrol specifically inhibited the motility of the mitotic kinesin Eg5, a motor protein required for spindle bipolarity, and will therefore be a particularly useful tool for studying mitotic mechanisms.
Journal ArticleDOI

Analysis of gene function in somatic mammalian cells using small interfering RNAs

TL;DR: Because of the robustness of the siRNA knockdown technology, genomewide analysis of human gene function in cultured cells has now become possible and here a collection of protocols for siRNA-mediated knockdown of mammalian gene expression is provided.
Journal ArticleDOI

Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement.

TL;DR: This article summarizes the current views on the dynamic structure of the mitotic spindle and its relation to mitotic chromosome movements based on measurements of birefringence of spindle fibers in living cells, normally developing or experimentally modified by various physical and chemical agents.
Journal ArticleDOI

Microtubule motors in mitosis

TL;DR: It is becoming clear that motors invoke several distinct mechanisms to generate the forces that drive mitosis, and in carrying out its function, the spindle appears to pass through a series of transient steady-state structures, each established by a delicate balance of forces generated by multiple complementary and antagonistic motors.
Journal ArticleDOI

Ran Induces Spindle Assembly by Reversing the Inhibitory Effect of Importin α on TPX2 Activity

TL;DR: It is shown that components of the nuclear transport machinery serve to regulate spindle formation in M phase, and TPX2 is required for Ran.GTP, a microtubule-associated protein previously known to target a motor protein, Xklp2, to microtubules.
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