Mechanisms of kinesin‐7 CENP‐E in kinetochore–microtubule capture and chromosome alignment during cell division

2019 ◽  
Vol 111 (6) ◽  
pp. 143-160 ◽  
Author(s):  
Kai‐Wei Yu ◽  
Ning Zhong ◽  
Yu Xiao ◽  
Zhen‐Yu She
Keyword(s):  
Cell Division ◽  
Chromosome Alignment ◽  
Microtubule Capture

scholarly journals Wnt signaling recruits KIF2A to the spindle to ensure chromosome congression and alignment during mitosis

2021 ◽  
Vol 118 (34) ◽  
pp. e2108145118
Author(s):  
Anja Bufe ◽  
Ana García del Arco ◽  
Magdalena Hennecke ◽  
Anchel de Jaime-Soguero ◽  
Matthias Ostermaier ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Division ◽  
Wnt Signaling ◽  
Pluripotent Stem Cells ◽  
Target Genes ◽  
Genome Maintenance ◽  
Spindle Poles ◽  
Chromosome Congression ◽  
Chromosome Alignment ◽  
Canonical Wnt

Canonical Wnt signaling plays critical roles in development and tissue renewal by regulating β-catenin target genes. Recent evidence showed that β-catenin–independent Wnt signaling is also required for faithful execution of mitosis. However, the targets and specific functions of mitotic Wnt signaling still remain uncharacterized. Using phosphoproteomics, we identified that Wnt signaling regulates the microtubule depolymerase KIF2A during mitosis. We found that Dishevelled recruits KIF2A via its N-terminal and motor domains, which is further promoted upon LRP6 signalosome formation during cell division. We show that Wnt signaling modulates KIF2A interaction with PLK1, which is critical for KIF2A localization at the spindle. Accordingly, inhibition of basal Wnt signaling leads to chromosome misalignment in somatic cells and pluripotent stem cells. We propose that Wnt signaling monitors KIF2A activity at the spindle poles during mitosis to ensure timely chromosome alignment. Our findings highlight a function of Wnt signaling during cell division, which could have important implications for genome maintenance, notably in stem cells.

scholarly journals Separase cleaves the kinetochore protein Meikin to direct the meiosis I/II transition

2020 ◽  
Author(s):  
Nolan K Maier ◽  
Jun Ma ◽  
Michael A Lampson ◽  
Iain M Cheeseman
Keyword(s):  
Cell Division ◽  
Germ Cells ◽  
Molecular Switch ◽  
Specific Protein ◽  
Cleavage Product ◽  
Regulatory Mechanisms ◽  
Kinetochore Protein ◽  
Chromosome Alignment ◽  
Activity State ◽  
Meiosis I

SummaryTo generate haploid gametes, germ cells undergo two consecutive meiotic divisions requiring key changes to the cell division machinery. Here, we explore the regulatory mechanisms that differentially control meiotic events. We demonstrate that the protease Separase rewires key cell division processes at the meiosis I/II transition by cleaving the meiosis-specific protein Meikin. In contrast to cohesin, which is inactivated by Separase proteolysis, cleaved Meikin remains functional, but results in a distinct activity state. Full-length Meikin and the C-terminal Meikin Separase-cleavage product both localize to kinetochores, bind to Plk1 kinase, and promote Rec8 cleavage, but our results reveal distinct roles for these proteins in controlling meiosis. Mutations that prevent Meikin cleavage or that conditionally inactivate Meikin at anaphase I both result in defective meiosis II chromosome alignment. Thus, Separase cleavage of Meikin creates an irreversible molecular switch to rewire the cell division machinery at the meiosis I/II transition.

scholarly journals Plasmodium Kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission

2019 ◽  
Author(s):  
Mohammad Zeeshan ◽  
Fiona Shilliday ◽  
Tianyang Liu ◽  
Steven Abel ◽  
Tobias Mourier ◽  
...  
Keyword(s):  
Life Cycle ◽  
Cell Division ◽  
Gene Targeting ◽  
Intracellular Transport ◽  
Regulation Of Gene Expression ◽  
Microtubule Depolymerization ◽  
Expression Of Genes ◽  
Genome Wide ◽  
Spindle Dynamics ◽  
Chromosome Alignment

AbstractKinesin-8 proteins are microtubule motors that are often involved in regulation of mitotic spindle length and chromosome alignment. They move towards the ends of spindle microtubules and regulate the dynamics of these ends due, at least in some species, to their microtubule depolymerization activity. Plasmodium spp. exhibit an atypical endomitotic cell division in which chromosome condensation and spindle dynamics are not well understood in the different proliferative stages. Genome-wide homology analysis of Plasmodium spp. revealed the presence of two Kinesin-8 motor proteins (Kinesin-8X and Kinesin-8B). Here we have studied the biochemical properties of Kinesin-8X and its role in parasite proliferation. In vitro, Kinesin-8X showed motile and depolymerization activities like other Kinesin-8 motors. To understand its role in cell division, we have used protein tagging and live cell imaging to define the location of Plasmodium Kinesin-8X during all proliferative stages of the P berghei life cycle. Furthermore, we have used gene targeting to analyse the function of Kinesin-8X. The results reveal a spatio-temporal involvement of Kinesin-8X in spindle dynamics and its association with both mitotic and meiotic spindles and the putative microtubule organising centre (MTOC). Deletion of the Kinesin-8X gene showed that this protein is required for endomitotic division during oocyst development and is therefore necessary for parasite replication within the mosquito gut, and for transmission to the vertebrate host. Consistently, transcriptome analysis of Δkinesin-8X parasites reveals modulated expression of genes involved mainly in microtubule-based processes, chromosome organisation and the regulation of gene expression supporting a role in cell division.Author SummaryKinesins are microtubule-based motors that play key roles in intracellular transport, cell division and motility. Members of the Kinesin-8 family contribute to chromosome alignment during cell division in many eukaryotes. However, the roles of kinesins in the atypical cell division in Plasmodium, the causative agent of malaria, is not known. In contrast to many other eukaryotes, Plasmodium proliferates by endomitosis, in which genome replication and division occur within a nucleus bounded by a persistent nuclear envelope. We show that the Plasmodium genome encodes only nine kinesins and we further investigate the role of Kinesin-8X throughout the Plasmodium life cycle using biochemical and gene targeting approaches. We show that Plasmodium Kinesin-8X has microtubule-based motility and depolymerization activity. We also show that Kinesin-8X is probably localized on putative MTOCs and spindles during cell division in most of the stages of P. berghei life cycle. By gene deletion we demonstrate that Kinesin-8X is essential for normal oocyst development and sporozoite formation. Genome-wide RNA analysis of Δkinesin-8X parasites reveals modulated expression of genes involved in microtubule-based processes. Overall, the data suggest that Kinesin-8X is a molecular motor that plays essential roles during endomitosis in oocyst development in the mosquito, contributing to parasite transmission.

scholarly journals The C-terminal helix of BubR1 is essential for CENP-E-dependent chromosome alignment

2020 ◽  
Author(s):  
Thibault Legal ◽  
Daniel Hayward ◽  
Agata Gluszek-Kustusz ◽  
Elizabeth A. Blackburn ◽  
Christos Spanos ◽  
...  
Keyword(s):  
Cell Division ◽  
Molecular Basis ◽  
Spindle Checkpoint ◽  
Checkpoint Protein ◽  
Molecular Determinants ◽  
Chromosome Alignment

AbstractDuring cell division, misaligned chromosomes are captured and aligned by motors before their segregation. The CENP-E motor is recruited to polar unattached kinetochores, to facilitate chromosome alignment. The spindle checkpoint protein BubR1 has been reported as a CENP-E interacting partner, but to what extent, if at all, BubR1 contributes to CENP-E localization at kinetochores, has remained controversial. Here we define the molecular determinants that specify the interaction between BubR1 and CENP-E. The basic C-terminal helix of BubR1 is necessary but not sufficient for CENP-E interaction, while a minimal key acidic patch on the kinetochore-targeting domain of CENP-E, is also essential. We then demonstrate that BubR1 is required for the recruitment of CENP-E to kinetochores to facilitate chromosome alignment. This BubR1-CENP-E axis is critical to align chromosomes that have failed to congress through other pathways and recapitulates the major known function of CENP-E. Overall, our studies define the molecular basis and the function for CENP-E recruitment to BubR1 at kinetochores during mammalian mitosis.

scholarly journals The pseudo GTPase CENP-M drives human kinetochore assembly

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Federica Basilico ◽  
Stefano Maffini ◽  
John R Weir ◽  
Daniel Prumbaum ◽  
Ana M Rojas ◽  
...  
Keyword(s):  
Cell Division ◽  
Small Gtpases ◽  
Conformational Switching ◽  
Point Mutant ◽  
Kinetochore Assembly ◽  
Gtp Binding ◽  
Chromosome Alignment ◽  
Spindle Microtubules ◽  
And Function

Kinetochores, multi-subunit complexes that assemble at the interface with centromeres, bind spindle microtubules to ensure faithful delivery of chromosomes during cell division. The configuration and function of the kinetochore–centromere interface is poorly understood. We report that a protein at this interface, CENP-M, is structurally and evolutionarily related to small GTPases but is incapable of GTP-binding and conformational switching. We show that CENP-M is crucially required for the assembly and stability of a tetramer also comprising CENP-I, CENP-H, and CENP-K, the HIKM complex, which we extensively characterize through a combination of structural, biochemical, and cell biological approaches. A point mutant affecting the CENP-M/CENP-I interaction hampers kinetochore assembly and chromosome alignment and prevents kinetochore recruitment of the CENP-T/W complex, questioning a role of CENP-T/W as founder of an independent axis of kinetochore assembly. Our studies identify a single pathway having CENP-C as founder, and CENP-H/I/K/M and CENP-T/W as CENP-C-dependent followers.

scholarly journals CENP-E–dependent BubR1 autophosphorylation enhances chromosome alignment and the mitotic checkpoint

2012 ◽  
Vol 198 (2) ◽  
pp. 205-217 ◽  
Author(s):  
Yige Guo ◽  
Christine Kim ◽  
Sana Ahmad ◽  
Jiayin Zhang ◽  
Yinghui Mao
Keyword(s):  
Mitotic Checkpoint ◽  
The State ◽  
Chromosome Loss ◽  
Spindle Microtubule ◽  
Single Chromosome ◽  
Checkpoint Signaling ◽  
Mitotic Kinase ◽  
Chromosome Alignment ◽  
Kinetochore Function ◽  
Microtubule Capture

How the state of spindle microtubule capture at the kinetochore is translated into mitotic checkpoint signaling remains largely unknown. In this paper, we demonstrate that the kinetochore-associated mitotic kinase BubR1 phosphorylates itself in human cells and that this autophosphorylation is dependent on its binding partner, the kinetochore motor CENP-E. This CENP-E–dependent BubR1 autophosphorylation at unattached kinetochores is important for a full-strength mitotic checkpoint to prevent single chromosome loss. Replacing endogenous BubR1 with a nonphosphorylatable BubR1 mutant, as well as depletion of CENP-E, the BubR1 kinase activator, results in metaphase chromosome misalignment and a decrease of Aurora B–mediated Ndc80 phosphorylation at kinetochores. Furthermore, expressing a phosphomimetic BubR1 mutant substantially reduces the incidence of polar chromosomes in CENP-E–depleted cells. Thus, the state of CENP-E–dependent BubR1 autophosphorylation in response to spindle microtubule capture by CENP-E is important for kinetochore function in achieving accurate chromosome segregation.

scholarly journals A centriole's subdistal appendages: contributions to cell division, ciliogenesis and differentiation

Open Biology ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 200399
Author(s):  
Nicole A. Hall ◽  
Heidi Hehnly
Keyword(s):  
Cell Division ◽  
Cell Differentiation ◽  
Final Stage ◽  
Eukaryotic Species ◽  
Chromosome Alignment ◽  
Cilia Formation ◽  
Mother Centriole ◽  
Pericentriolar Material ◽  
Daughter Centriole

The centrosome is a highly conserved structure composed of two centrioles surrounded by pericentriolar material. The mother, and inherently older, centriole has distal and subdistal appendages, whereas the daughter centriole is devoid of these appendage structures. Both appendages have been primarily linked to functions in cilia formation. However, subdistal appendages present with a variety of potential functions that include spindle placement, chromosome alignment, the final stage of cell division (abscission) and potentially cell differentiation. Subdistal appendages are particularly interesting in that they do not always display a conserved ninefold symmetry in appendage organization on the mother centriole across eukaryotic species, unlike distal appendages. In this review, we aim to differentiate both the morphology and role of the distal and subdistal appendages, with a particular focus on subdistal appendages.

scholarly journals The chromosomal passenger complex: guiding Aurora-B through mitosis

2006 ◽  
Vol 173 (6) ◽  
pp. 833-837 ◽  
Author(s):  
Gerben Vader ◽  
René H. Medema ◽  
Susanne M.A. Lens
Keyword(s):  
Cell Division ◽  
Histone Modification ◽  
Posttranslational Modifications ◽  
Molecular Mechanisms ◽  
Aurora B ◽  
Chromosomal Passenger Complex ◽  
Precise Control ◽  
Chromosome Alignment ◽  
Chromosomal Passenger ◽  
And Function

During mitosis, the chromosomal passenger complex (CPC) orchestrates highly different processes, such as chromosome alignment, histone modification, and cytokinesis. Proper and timely localization of this complex is the key to precise control over the enzymatic core of the CPC, the Aurora-B kinase. We discuss the molecular mechanisms by which the CPC members direct the dynamic localization of the complex throughout cell division. Also, we summarize posttranslational modifications that occur on the CPC and discuss their roles in regulating localization and function of this mitotic complex.

scholarly journals The C-terminal helix of BubR1 is essential for CENP-E-dependent chromosome alignment

2020 ◽  
Vol 133 (16) ◽  
pp. jcs246025 ◽  
Author(s):  
Thibault Legal ◽  
Daniel Hayward ◽  
Agata Gluszek-Kustusz ◽  
Elizabeth A. Blackburn ◽  
Christos Spanos ◽  
...  
Keyword(s):  
Cell Division ◽  
Molecular Basis ◽  
Spindle Checkpoint ◽  
First Person ◽  
Checkpoint Protein ◽  
Molecular Determinants ◽  
Chromosome Alignment

ABSTRACTDuring cell division, misaligned chromosomes are captured and aligned by motors before their segregation. The CENP-E motor is recruited to polar unattached kinetochores to facilitate chromosome alignment. The spindle checkpoint protein BubR1 (also known as BUB1B) has been reported as a CENP-E interacting partner, but the extent to which BubR1 contributes to CENP-E localization at kinetochores has remained controversial. Here we define the molecular determinants that specify the interaction between BubR1 and CENP-E. The basic C-terminal helix of BubR1 is necessary but not sufficient for CENP-E interaction, and a minimal key acidic patch on the kinetochore-targeting domain of CENP-E is also essential. We then demonstrate that BubR1 is required for the recruitment of CENP-E to kinetochores to facilitate chromosome alignment. This BubR1–CENP-E axis is critical for alignment of chromosomes that have failed to congress through other pathways and recapitulates the major known function of CENP-E. Overall, our studies define the molecular basis and the function for CENP-E recruitment to BubR1 at kinetochores during mammalian mitosis.This article has an associated First Person interview with the first author of the paper.

scholarly journals Mitotic chromosome alignment ensures mitotic fidelity by promoting interchromosomal compaction during anaphase

2019 ◽  
Vol 218 (4) ◽  
pp. 1148-1163 ◽  
Author(s):  
Cindy L. Fonseca ◽  
Heidi L.H. Malaby ◽  
Leslie A. Sepaniac ◽  
Whitney Martin ◽  
Candice Byers ◽  
...  
Keyword(s):  
Cell Division ◽  
Postnatal Growth ◽  
Mitotic Chromosomes ◽  
Growth And Survival ◽  
Copy Numbers ◽  
Chromosome Alignment ◽  
Single Nucleus ◽  
Complete Cell

Chromosome alignment at the equator of the mitotic spindle is a highly conserved step during cell division; however, its importance to genomic stability and cellular fitness is not understood. Normal mammalian somatic cells lacking KIF18A function complete cell division without aligning chromosomes. These alignment-deficient cells display normal chromosome copy numbers in vitro and in vivo, suggesting that chromosome alignment is largely dispensable for maintenance of euploidy. However, we find that loss of chromosome alignment leads to interchromosomal compaction defects during anaphase, abnormal organization of chromosomes into a single nucleus at mitotic exit, and the formation of micronuclei in vitro and in vivo. These defects slow cell proliferation and are associated with impaired postnatal growth and survival in mice. Our studies support a model in which the alignment of mitotic chromosomes promotes proper organization of chromosomes into a single nucleus and continued proliferation by ensuring that chromosomes segregate as a compact mass during anaphase.

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