scholarly journals Reductionism at the vertebrate kinetochore

2012 ◽  
Vol 200 (1) ◽  
pp. 7-8 ◽  
Author(s):  
Ana Stankovic ◽  
Lars E.T. Jansen
Keyword(s):  
Chromosome Segregation ◽  
Protein Interactions ◽  
Mitotic Spindle ◽  
De Novo ◽  
Large Structure ◽  
Cell Biol ◽  
Spindle Microtubules ◽  
The Creation

The kinetochore forms the site of attachment for mitotic spindle microtubules driving chromosome segregation. The interdependent protein interactions in this large structure have made it difficult to dissect the function of its components. In this issue, Hori et al. (2013. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201210106) present a novel and powerful methodology to address the sufficiency of individual proteins for the creation of a functional de novo centromere.

scholarly journals Concentrating on the mitotic spindle

2015 ◽  
Vol 210 (5) ◽  
pp. 691-693 ◽  
Author(s):  
Paul S. Maddox ◽  
Anne-Marie Ladouceur
Keyword(s):  
Chromosome Segregation ◽  
Mitotic Spindle ◽  
Molecular Crowding ◽  
Cell Biol ◽  
New Feature

In eukaryotes, the microtubule-based spindle drives chromosome segregation. In this issue, Schweizer et al. (2015; J. Cell Biol. http://dx.doi.org/10.1083/jcb.201506107) find that the spindle area is demarcated by a semipermeable organelle barrier. Molecular crowding, which is microtubule independent, causes the enrichment and/or retention of crucial factors in the spindle region. Their results add an important new feature to the models of how this structure assembles and is regulated.

scholarly journals The budding yeast Ipl1/Aurora protein kinase regulates mitotic spindle disassembly

2003 ◽  
Vol 160 (3) ◽  
pp. 329-339 ◽  
Author(s):  
Stéphanie Buvelot ◽  
Sean Y. Tatsutani ◽  
Danielle Vermaak ◽  
Sue Biggins
Keyword(s):  
Chromosome Segregation ◽  
Mitotic Spindle ◽  
Kinase Activity ◽  
Budding Yeast ◽  
Genomic Stability ◽  
Time Lapse ◽  
Time Lapse Microscopy ◽  
Spindle Disassembly ◽  
Spindle Midzone ◽  
Spindle Microtubules

Ipl1p is the budding yeast member of the Aurora family of protein kinases, critical regulators of genomic stability that are required for chromosome segregation, the spindle checkpoint, and cytokinesis. Using time-lapse microscopy, we found that Ipl1p also has a function in mitotic spindle disassembly that is separable from its previously identified roles. Ipl1–GFP localizes to kinetochores from G1 to metaphase, transfers to the spindle after metaphase, and accumulates at the spindle midzone late in anaphase. Ipl1p kinase activity increases at anaphase, and ipl1 mutants can stabilize fragile spindles. As the spindle disassembles, Ipl1p follows the plus ends of the depolymerizing spindle microtubules. Many Ipl1p substrates colocalize with Ipl1p to the spindle midzone, identifying additional proteins that may regulate spindle disassembly. We propose that Ipl1p regulates both the kinetochore and interpolar microtubule plus ends to regulate its various mitotic functions.

scholarly journals Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly.

1992 ◽  
Vol 118 (1) ◽  
pp. 109-120 ◽  
Author(s):  
M A Hoyt ◽  
L He ◽  
K K Loo ◽  
W S Saunders
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Viability ◽  
Chromosome Segregation ◽  
Heavy Chain ◽  
Mitotic Spindle ◽  
Spindle Assembly ◽  
Gene Products ◽  
Related Gene ◽  
Mitotic Spindle Assembly ◽  
Spindle Microtubules

Two Saccharomyces cerevisiae genes, CIN8 and KIP1 (a.k.a. CIN9), were identified by their requirement for normal chromosome segregation. Both genes encode polypeptides related to the heavy chain of the microtubule-based force-generating enzyme kinesin. Cin8p was found to be required for pole separation during mitotic spindle assembly at 37 degrees C, although overproduced Kip1p could substitute. At lower temperatures, the activity of at least one of these proteins was required for cell viability, indicating that they perform an essential but redundant function. Cin8p was observed to be a component of the mitotic spindle, colocalizing with the microtubules that lie between the poles. Taken together, these findings suggest that these proteins interact with spindle microtubules to produce an outwardly directed force acting upon the poles.

scholarly journals Aurora at the pole and equator: overlapping functions of Aurora kinases in the mitotic spindle

Open Biology ◽  
2013 ◽  
Vol 3 (3) ◽  
pp. 120185 ◽  
Author(s):  
Helfrid Hochegger ◽  
Nadia Hégarat ◽  
Jose B. Pereira-Leal
Keyword(s):  
Chromosome Segregation ◽  
Mitotic Spindle ◽  
Aurora Kinase ◽  
Spindle Pole ◽  
Aurora A ◽  
Mitotic Progression ◽  
Aurora Kinases ◽  
Mitotic Kinases ◽  
Chromosome Alignment ◽  
Spindle Microtubules

The correct assembly and timely disassembly of the mitotic spindle is crucial for the propagation of the genome during cell division. Aurora kinases play a central role in orchestrating bipolar spindle establishment, chromosome alignment and segregation. In most eukaryotes, ranging from amoebas to humans, Aurora activity appears to be required both at the spindle pole and the kinetochore, and these activities are often split between two different Aurora paralogues, termed Aurora A and B. Polar and equatorial functions of Aurora kinases have generally been considered separately, with Aurora A being mostly involved in centrosome dynamics, whereas Aurora B coordinates kinetochore attachment and cytokinesis. However, double inactivation of both Aurora A and B results in a dramatic synergy that abolishes chromosome segregation. This suggests that these two activities jointly coordinate mitotic progression. Accordingly, recent evidence suggests that Aurora A and B work together in both spindle assembly in metaphase and disassembly in anaphase. Here, we provide an outlook on these shared functions of the Auroras, discuss the evolution of this family of mitotic kinases and speculate why Aurora kinase activity may be required at both ends of the spindle microtubules.

scholarly journals Cytokinetic astralogy

2009 ◽  
Vol 187 (6) ◽  
pp. 757-759 ◽  
Author(s):  
Julie C. Canman
Keyword(s):  
Mitotic Spindle ◽  
Direct Contact ◽  
Cell Cortex ◽  
Animal Cells ◽  
Division Plane ◽  
Cell Biol ◽  
Spindle Microtubules

Division plane specification in animal cells has long been presumed to involve direct contact between microtubules of the anaphase mitotic spindle and the cell cortex. In this issue, von Dassow et al. (von Dassow et al. 2009. J. Cell. Biol. doi:10.1083/jcb.200907090) challenge this assumption by showing that spindle microtubules can effectively position the division plane at a distance from the cell cortex.

scholarly journals Separate to operate: control of centrosome positioning and separation

2014 ◽  
Vol 369 (1650) ◽  
pp. 20130461 ◽  
Author(s):  
Fikret G. Agircan ◽  
Elmar Schiebel ◽  
Balca R. Mardin
Keyword(s):  
Cell Cycle ◽  
Cell Motility ◽  
Cell Polarity ◽  
Chromosome Segregation ◽  
Mitotic Spindle ◽  
De Novo Assembly ◽  
De Novo ◽  
Animal Cells ◽  
Pericentriolar Material ◽  
Golgi Organization

The centrosome is the main microtubule (MT)-organizing centre of animal cells. It consists of two centrioles and a multi-layered proteinaceous structure that surrounds the centrioles, the so-called pericentriolar material. Centrosomes promote de novo assembly of MTs and thus play important roles in Golgi organization, cell polarity, cell motility and the organization of the mitotic spindle. To execute these functions, centrosomes have to adopt particular cellular positions. Actin and MT networks and the association of the centrosomes to the nuclear envelope define the correct positioning of the centrosomes. Another important feature of centrosomes is the centrosomal linker that connects the two centrosomes. The centrosome linker assembles in late mitosis/G1 simultaneously with centriole disengagement and is dissolved before or at the beginning of mitosis. Linker dissolution is important for mitotic spindle formation, and its cell cycle timing has profound influences on the execution of mitosis and proficiency of chromosome segregation. In this review, we will focus on the mechanisms of centrosome positioning and separation, and describe their functions and mechanisms in the light of recent findings.

scholarly journals A new piece in the kinetochore jigsaw puzzle

2014 ◽  
Vol 206 (4) ◽  
pp. 457-459
Author(s):  
Kevin D. Corbett ◽  
Arshad Desai
Keyword(s):  
Cell Division ◽  
Signaling Pathways ◽  
Chromosome Segregation ◽  
Budding Yeast ◽  
Eukaryotic Cell ◽  
Jigsaw Puzzle ◽  
New Model ◽  
Cell Biol ◽  
Chromosome Attachment ◽  
Spindle Microtubules

In eukaryotic cell division, the kinetochore mediates chromosome attachment to spindle microtubules and acts as a scaffold for signaling pathways, ensuring the accuracy of chromosome segregation. The architecture of the kinetochore underlies its function in mitosis. In this issue, Hornung et al. (2014. J. Cell Biol. http://dx.doi.org/201403081) identify an unexpected linkage between the inner and outer regions of the kinetochore in budding yeast that suggests a new model for the construction of this interface.

scholarly journals De Novo Kinetochore Assembly Requires the Centromeric Histone H3 Variant

2005 ◽  
Vol 16 (12) ◽  
pp. 5649-5660 ◽  
Author(s):  
Kimberly A. Collins ◽  
Andrea R. Castillo ◽  
Sean Y. Tatsutani ◽  
Sue Biggins
Keyword(s):  
Chromosome Segregation ◽  
Mitotic Spindle ◽  
De Novo ◽  
Histone H3 ◽  
Centromeric Chromatin ◽  
Centromeric Dna ◽  
Kinetochore Assembly ◽  
Histone H3 Variant ◽  
Chromosome Attachment ◽  
Dna Specificity

Kinetochores mediate chromosome attachment to the mitotic spindle to ensure accurate chromosome segregation. Budding yeast is an excellent organism for kinetochore assembly studies because it has a simple defined centromere sequence responsible for the localization of >65 proteins. In addition, yeast is the only organism where a conditional centromere is available to allow studies of de novo kinetochore assembly. Using a conditional centromere, we found that yeast kinetochore assembly is not temporally restricted and can occur in both G1 phase and prometaphase. We performed the first investigation of kinetochore assembly in the absence of the centromeric histone H3 variant Cse4 and found that all proteins tested depend on Cse4 to localize. Consistent with this observation, Cse4-depleted cells had severe chromosome segregation defects. We therefore propose that yeast kinetochore assembly requires both centromeric DNA specificity and centromeric chromatin.

scholarly journals An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast

2018 ◽  
Author(s):  
Jackie Lang ◽  
Adrienne Barber ◽  
Sue Biggins
Keyword(s):  
Cell Cycle ◽  
Chromosome Segregation ◽  
De Novo ◽  
Budding Yeast ◽  
Centromeric Dna ◽  
Kinetochore Assembly ◽  
Ndc80 Complex ◽  
Spindle Microtubules ◽  
Underlying Mechanisms ◽  
Mitotic Phosphorylation

ABSTRACTChromosome segregation depends on the kinetochore, the machine that establishes force-bearing attachments between DNA and spindle microtubules. Kinetochores are formed every cell cycle via a highly regulated process that requires coordinated assembly of multiple subcomplexes on specialized chromatin. To elucidate the underlying mechanisms, we developed an assay to assemble kinetochores de novo using centromeric DNA and budding yeast extracts. Assembly is enhanced by mitotic phosphorylation of the Dsn1 kinetochore protein and generates kinetochores capable of binding microtubules. We used this assay to investigate why kinetochores recruit the microtubule-binding Ndc80 complex via two receptors: the Mis12 complex and CENP-T. Although the CENP-T pathway is non-essential in yeast, we demonstrate that it becomes essential for viability and Ndc80c recruitment when the Mis12 pathway is crippled by defects in Dsn1 phosphorylation. Assembling kinetochores de novo in yeast extracts provides a powerful and genetically tractable method to elucidate critical regulatory events in the future.

scholarly journals An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Jackie Lang ◽  
Adrienne Barber ◽  
Sue Biggins
Keyword(s):  
Cell Cycle ◽  
Chromosome Segregation ◽  
De Novo ◽  
Budding Yeast ◽  
Centromeric Dna ◽  
Kinetochore Assembly ◽  
Ndc80 Complex ◽  
Spindle Microtubules ◽  
Underlying Mechanisms ◽  
Mitotic Phosphorylation

Chromosome segregation depends on the kinetochore, the machine that establishes force-bearing attachments between DNA and spindle microtubules. Kinetochores are formed every cell cycle via a highly regulated process that requires coordinated assembly of multiple subcomplexes on specialized chromatin. To elucidate the underlying mechanisms, we developed an assay to assemble kinetochores de novo using centromeric DNA and budding yeast extracts. Assembly is enhanced by mitotic phosphorylation of the Dsn1 kinetochore protein and generates kinetochores capable of binding microtubules. We used this assay to investigate why kinetochores recruit the microtubule-binding Ndc80 complex via two receptors: the Mis12 complex and CENP-T. Although the CENP-T pathway is non-essential in yeast, we demonstrate that it becomes essential for viability and Ndc80c recruitment when the Mis12 pathway is crippled by defects in Dsn1 phosphorylation. Assembling kinetochores de novo in yeast extracts provides a powerful and genetically tractable method to elucidate critical regulatory events in the future.

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