scholarly journals Kinesin-5 promotes microtubule nucleation and assembly by stabilizing a lattice-competent conformation of tubulin

2019 ◽  
Author(s):  
Geng-Yuan Chen ◽  
Ana B. Asenjo ◽  
Yalei Chen ◽  
Jake Mascaro ◽  
David F. J. Arginteanu ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Polymerase Activity ◽  
Microtubule Nucleation ◽  
Domain Swap ◽  
Microtubule Polymerization ◽  
The Family ◽  
Microtubule Growth ◽  
Spindle Elongation ◽  
Necessary And Sufficient ◽  
Induced Changes

SummaryBesides sliding apart antiparallel microtubules during spindle elongation, the mitotic kinesin-5, Eg5 promotes microtubule polymerization, emphasizing its importance in mitotic spindle length control. Here, we characterize the Eg5 microtubule polymerase mechanism by assessing motor-induced changes in the longitudinal and lateral tubulin-tubulin bonds that form the microtubule lattice. Isolated Eg5 motor domains promote microtubule nucleation, growth and stability. Eg5 binds preferentially to microtubules over free tubulin, and colchicine-like inhibitors that stabilize the bent conformation of tubulin allosterically inhibit Eg5 binding, consistent with a model in which Eg5 induces a curved-to-straight transition in tubulin. Domain swap experiments establish that the family-specific Loop11, which resides near the nucleotide-sensing Switch-II domain, is necessary and sufficient for the polymerase activity of Eg5. Thus, we propose a microtubule polymerase mechanism in which Eg5 at the plus-end promotes a curved-to-straight transition in tubulin that enhances lateral bond formation and thereby promotes microtubule growth and stability.

scholarly journals The kinesin-8 Kip3 scales anaphase spindle length by suppression of midzone microtubule polymerization

2014 ◽  
Vol 204 (6) ◽  
pp. 965-975 ◽  
Author(s):  
Rania S. Rizk ◽  
Katherine A. DiScipio ◽  
Kathleen G. Proudfoot ◽  
Mohan L. Gupta
Keyword(s):  
Mitotic Spindle ◽  
Molecular Mechanisms ◽  
Spindle Microtubule ◽  
Microtubule Polymerization ◽  
Sister Chromatids ◽  
Final Length ◽  
Spindle Microtubules ◽  
Spindle Elongation ◽  
Spindle Function ◽  
Yeast Mutants

Mitotic spindle function is critical for cell division and genomic stability. During anaphase, the elongating spindle physically segregates the sister chromatids. However, the molecular mechanisms that determine the extent of anaphase spindle elongation remain largely unclear. In a screen of yeast mutants with altered spindle length, we identified the kinesin-8 Kip3 as essential to scale spindle length with cell size. Kip3 is a multifunctional motor protein with microtubule depolymerase, plus-end motility, and antiparallel sliding activities. Here we demonstrate that the depolymerase activity is indispensable to control spindle length, whereas the motility and sliding activities are not sufficient. Furthermore, the microtubule-destabilizing activity is required to counteract Stu2/XMAP215-mediated microtubule polymerization so that spindle elongation terminates once spindles reach the appropriate final length. Our data support a model where Kip3 directly suppresses spindle microtubule polymerization, limiting midzone length. As a result, sliding forces within the midzone cannot buckle spindle microtubules, which allows the cell boundary to define the extent of spindle elongation.

scholarly journals Ensconsin/Map7 promotes microtubule growth and centrosome separation in Drosophila neural stem cells

2014 ◽  
Vol 204 (7) ◽  
pp. 1111-1121 ◽  
Author(s):  
Emmanuel Gallaud ◽  
Renaud Caous ◽  
Aude Pascal ◽  
Franck Bazile ◽  
Jean-Philippe Gagné ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Spindle Assembly ◽  
Microtubule Associated Proteins ◽  
Microtubule Polymerization ◽  
Sister Chromatids ◽  
Centrosome Separation ◽  
Associated Proteins ◽  
Microtubule Growth ◽  
Mitotic Spindle Assembly

The mitotic spindle is crucial to achieve segregation of sister chromatids. To identify new mitotic spindle assembly regulators, we isolated 855 microtubule-associated proteins (MAPs) from Drosophila melanogaster mitotic or interphasic embryos. Using RNAi, we screened 96 poorly characterized genes in the Drosophila central nervous system to establish their possible role during spindle assembly. We found that Ensconsin/MAP7 mutant neuroblasts display shorter metaphase spindles, a defect caused by a reduced microtubule polymerization rate and enhanced by centrosome ablation. In agreement with a direct effect in regulating spindle length, Ensconsin overexpression triggered an increase in spindle length in S2 cells, whereas purified Ensconsin stimulated microtubule polymerization in vitro. Interestingly, ensc-null mutant flies also display defective centrosome separation and positioning during interphase, a phenotype also detected in kinesin-1 mutants. Collectively, our results suggest that Ensconsin cooperates with its binding partner Kinesin-1 during interphase to trigger centrosome separation. In addition, Ensconsin promotes microtubule polymerization during mitosis to control spindle length independent of Kinesin-1.

scholarly journals Spindle scaling is governed by cell boundary regulation of microtubule nucleation

2020 ◽  
Author(s):  
Elisa Maria Rieckhoff ◽  
Frederic Berndt ◽  
Stefan Golfier ◽  
Franziska Decker ◽  
Maria Elsner ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Large Range ◽  
Zebrafish Embryos ◽  
Quantitative Microscopy ◽  
Microtubule Nucleation ◽  
Microtubule Polymerization ◽  
Egg Extracts ◽  
Boundary Regulation ◽  
Geometric Cues ◽  
Cellular Organelles

AbstractCellular organelles such as the mitotic spindle adjust their size to the dimensions of the cell. It is widely understood that spindle scaling is governed by regulation of microtubule polymerization. Here we use quantitative microscopy in living zebrafish embryos and Xenopus egg extracts in combination with theory to show that microtubule polymerization dynamics are insufficient to scale spindles and only contribute below a critical cell size. In contrast, microtubule nucleation governs spindle scaling for all cell sizes. We show that this hierarchical regulation arises from the partitioning of a nucleation inhibitor to the cell membrane. Our results reveal that cells differentially regulate microtubule number and length using distinct geometric cues to maintain a functional spindle architecture over a large range of cell sizes.

scholarly journals Microtubule associated proteins and motors required for ectopic microtubule array formation in S. cerevisiae

2021 ◽  
Author(s):  
Brianna R. King ◽  
Janet B. Meehl ◽  
Tamira Vojnar ◽  
Mark Winey ◽  
Eric G. Muller ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Polymerase Activity ◽  
Nucleation Site ◽  
Microtubule Nucleation ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
A Subunit ◽  
Nucleation Activity

AbstractThe mitotic spindle is resilient to perturbation due to the concerted, and sometimes redundant, action of motors and microtubule-associated proteins. Here we utilize an inducible ectopic microtubule nucleation site in the nucleus of Saccharomyces cerevisiae to study three necessary steps in the formation of a bipolar array: the recruitment of the γ-tubulin complex, nucleation and elongation of microtubules, and the organization of microtubules relative to each other. This novel tool, an Spc110 chimera, reveals previously unreported roles of the microtubule-associated proteins Stu2, Bim1, and Bik1, and the motors Vik1 and Kip3. We report that Stu2 and Bim1 are required for nucleation and that Bik1 and Kip3 promote nucleation at the ectopic site. Stu2, Bim1, and Kip3 join their homologs XMAP215, EB1 and kinesin-8 as promoters of microtubule nucleation, while Bik1 promotes MT nucleation indirectly via its role in SPB positioning. Further, we find that the nucleation activity of Stu2 in vivo correlates with its polymerase activity in vitro. Finally, we provide the first evidence that Vik1, a subunit of Kar3/Vik1 kinesin-14, promotes microtubule minus end focusing at the ectopic site.

scholarly journals TOG–tubulin binding specificity promotes microtubule dynamics and mitotic spindle formation

2017 ◽  
Vol 216 (6) ◽  
pp. 1641-1657 ◽  
Author(s):  
Amy E. Byrnes ◽  
Kevin C. Slep
Keyword(s):  
Mitotic Spindle ◽  
Spindle Assembly Checkpoint ◽  
Polymerase Activity ◽  
Microtubule Dynamics ◽  
Spindle Formation ◽  
Microtubule Binding ◽  
Microtubule Polymerization ◽  
Tubulin Binding ◽  
Specific Order ◽  
Spindle Function

XMAP215, CLASP, and Crescerin use arrayed tubulin-binding tumor overexpressed gene (TOG) domains to modulate microtubule dynamics. We hypothesized that TOGs have distinct architectures and tubulin-binding properties that underlie each family’s ability to promote microtubule polymerization or pause. As a model, we investigated the pentameric TOG array of a Drosophila melanogaster XMAP215 member, Msps. We found that Msps TOGs have distinct architectures that bind either free or polymerized tubulin, and that a polarized array drives microtubule polymerization. An engineered TOG1-2-5 array fully supported Msps-dependent microtubule polymerase activity. Requisite for this activity was a TOG5-specific N-terminal HEAT repeat that engaged microtubule lattice-incorporated tubulin. TOG5–microtubule binding maintained mitotic spindle formation as deleting or mutating TOG5 compromised spindle architecture and increased the mitotic index. Mad2 knockdown released the spindle assembly checkpoint triggered when TOG5–microtubule binding was compromised, indicating that TOG5 is essential for spindle function. Our results reveal a TOG5-specific role in mitotic fidelity and support our hypothesis that architecturally distinct TOGs arranged in a sequence-specific order underlie TOG array microtubule regulator activity.

The role of the augmin complex in establishing microtubule arrays

2019 ◽  
Vol 70 (12) ◽  
pp. 3035-3041
Author(s):  
Juan Tian ◽  
Zhaosheng Kong
Keyword(s):  
Cell Cycle ◽  
Plant Cells ◽  
Cell Type ◽  
Microtubule Nucleation ◽  
Microtubule Polymerization ◽  
Specific Composition ◽  
Ring Complex ◽  
Cell Type Specific ◽  
Microtubule Growth

Abstract Microtubule-dependent microtubule nucleation occurs on the lateral surface of pre-existing microtubules and provides a highly efficient means of amplifying their populations and reorganizing their architectures. The γ‑tubulin ring complex serves as the template to initiate nascent microtubule polymerization. Augmin, a hetero-octameric protein complex, acts as a recruiting factor to target the γ‑tubulin ring complex to pre-existing microtubules and trigger new microtubule growth. Although microtubule-dependent microtubule nucleation has been extensively studied in both animal and plant cells, it remains unclear how the augmin complex assembles in plant cells, especially in cell-cycle-specific and cell-type-specific manners, and how its spatial structure orchestrates the nucleation geometry. In this review, we summarize the advances in knowledge of augmin-dependent microtubule nucleation and the regulation of its geometry, and highlight recent findings and emerging questions concerning the role of the augmin complex in establishing microtubule arrays and the cell-cycle-specific composition of augmin in plant cells.

scholarly journals Microtubule associated proteins and motors required for ectopic microtubule array formation in S. cerevisiae

Genetics ◽  
2021 ◽  
Author(s):  
Brianna R King ◽  
Janet B Meehl ◽  
Tamira Vojnar ◽  
Mark Winey ◽  
Eric G Muller ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Polymerase Activity ◽  
Nucleation Site ◽  
Microtubule Nucleation ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
A Subunit ◽  
Nucleation Activity

Abstract The mitotic spindle is resilient to perturbation due to the concerted, and sometimes redundant, action of motors and microtubule-associated proteins. Here we utilize an inducible ectopic microtubule nucleation site in the nucleus of Saccharomyces cerevisiae to study three necessary steps in the formation of a bipolar array: the recruitment of the γ-tubulin complex, nucleation and elongation of microtubules, and the organization of microtubules relative to each other. This novel tool, an Spc110 chimera, reveals previously unreported roles of the microtubule-associated proteins Stu2, Bim1, and Bik1, and the motors Vik1 and Kip3. We report that Stu2 and Bim1 are required for nucleation and that Bik1 and Kip3 promote nucleation at the ectopic site. Stu2, Bim1, and Kip3 join their homologs XMAP215, EB1 and kinesin-8 as promoters of microtubule nucleation, while Bik1 promotes MT nucleation indirectly via its role in SPB positioning. Further, we find that the nucleation activity of Stu2 in vivo correlates with its polymerase activity in vitro. Finally, we provide the first evidence that Vik1, a subunit of Kar3/Vik1 kinesin-14, promotes microtubule minus end focusing at the ectopic site.

scholarly journals CAMSAP2 organizes a γ-tubulin-independent microtubule nucleation centre

2021 ◽  
Author(s):  
Tsuyoshi Imasaki ◽  
Satoshi Kikkawa ◽  
Shinsuke Niwa ◽  
Yumiko Saijo-Hamano ◽  
Hideki Shigematsu ◽  
...  
Keyword(s):  
Critical Concentration ◽  
Polymerization Process ◽  
Microtubule Nucleation ◽  
Nucleation Barrier ◽  
Microtubule Polymerization ◽  
Ring Complex ◽  
Nucleation Centre ◽  
Microtubule Formation ◽  
Microtubule Growth ◽  
In Cells

Microtubules are dynamic polymers consisting of αβ-tubulin heterodimers. The initial polymerization process, called microtubule nucleation, occurs spontaneously via αβ-tubulin. Since a large energy barrier prevents microtubule nucleation in cells, the γ-tubulin ring complex is recruited to the centrosome to overcome the nucleation barrier. However, detachment of a considerable number of microtubules from the centrosome is known to contribute to fundamental processes in cells. Here, we present evidence that minus-end-binding calmodulin-regulated spectrin-associated protein 2 (CAMSAP2) serves as a strong nucleator for microtubule formation from soluble αβ-tubulin independent of γ-tubulin. CAMSAP2 significantly reduces the nucleation barrier close to the critical concentration for microtubule polymerization by stabilizing the longitudinal contacts among αβ-tubulins. CAMSAP2 clusters together with αβ-tubulin to generate nucleation intermediates, from which numerous microtubules radiate, forming aster-like structures. Our findings suggest that CAMSAP2 supports microtubule growth by organizing a nucleation centre as well as by stabilizing microtubule nucleation intermediates.

Using binary operations to construct a transitive set of block transformations

2020 ◽  
Vol 30 (6) ◽  
pp. 375-389
Author(s):  
Igor V. Cherednik
Keyword(s):  
Binary Operation ◽  
Sufficient Conditions ◽  
Necessary And Sufficient Conditions ◽  
Binary Operations ◽  
The Family ◽  
Transitive Set ◽  
Necessary And Sufficient

AbstractWe study the set of transformations {ΣF : F∈ 𝓑∗(Ω)} implemented by a network Σ with a single binary operation F, where 𝓑∗(Ω) is the set of all binary operations on Ω that are invertible as function of the second variable. We state a criterion of bijectivity of all transformations from the family {ΣF : F∈ 𝓑∗(Ω)} in terms of the structure of the network Σ, identify necessary and sufficient conditions of transitivity of the set of transformations {ΣF : F∈ 𝓑∗(Ω)}, and propose an efficient way of verifying these conditions. We also describe an algorithm for construction of networks Σ with transitive sets of transformations {ΣF : F∈ 𝓑∗(Ω)}.

Binomial subsampling

2006 ◽  
Vol 462 (2068) ◽  
pp. 1181-1195 ◽  
Author(s):  
Carsten Wiuf ◽  
Michael P.H Stumpf
Keyword(s):  
Mathematical Biology ◽  
Power Series ◽  
Generating Functions ◽  
Binomial Distribution ◽  
Random Variable ◽  
Sufficient Condition ◽  
Necessary And Sufficient Condition ◽  
The Family ◽  
Necessary And Sufficient ◽  
Finite Moments

In this paper, we discuss statistical families with the property that if the distribution of a random variable X is in , then so is the distribution of Z ∼Bi( X ,  p ) for 0≤ p ≤1. (Here we take Z ∼Bi( X ,  p ) to mean that given X = x ,  Z is a draw from the binomial distribution Bi( x ,  p ).) It is said that the family is closed under binomial subsampling. We characterize such families in terms of probability generating functions and for families with finite moments of all orders we give a necessary and sufficient condition for the family to be closed under binomial subsampling. The results are illustrated with power series and other examples, and related to examples from mathematical biology. Finally, some issues concerning inference are discussed.

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