Effects of the microtubule nucleator Mto1 on chromosomal movement, DNA repair, and sister chromatid cohesion in fission yeast

Although the function of microtubules (MTs) in chromosomal segregation during mitosis is well characterized, much less is known about the role of MTs in chromosomal functions during interphase. In the fission yeast Schizosaccharomyces pombe, dynamic cytoplasmic MT bundles move chromosomes in an oscillatory manner during interphase via linkages through the nuclear envelope (NE) at the spindle pole body (SPB) and other sites. Mto1 is a cytoplasmic factor that mediates the nucleation and attachment of cytoplasmic MTs to the nucleus. Here, we test the function of these cytoplasmic MTs and Mto1 on DNA repair and recombination during interphase. We find that mto1Δ cells exhibit defects in DNA repair and homologous recombination (HR) and abnormal DNA repair factory dynamics. In these cells, sister chromatids are not properly paired, and binding of Rad21 cohesin subunit along chromosomal arms is reduced. Our findings suggest a model in which cytoplasmic MTs and Mto1 facilitate efficient DNA repair and HR by promoting dynamic chromosomal organization and cohesion in the nucleus.

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Centrosomal Aki1 and cohesin function in separase-regulated centriole disengagement

The Journal of Cell Biology ◽

10.1083/jcb.200906019 ◽

2009 ◽

Vol 187 (5) ◽

pp. 607-614 ◽

Cited By ~ 59

Author(s):

Akito Nakamura ◽

Hiroyuki Arai ◽

Naoya Fujita

Keyword(s):

Molecular Mechanism ◽

Sister Chromatid ◽

Dual Role ◽

Interacting Protein ◽

Concurrent Control ◽

Sister Chromatids ◽

Multipolar Spindles ◽

Chromatid Cohesion ◽

Cohesin Subunit

Sister chromatid separation at anaphase is triggered by cleavage of the cohesin subunit Scc1, which is mediated by separase. Centriole disengagement also requires separase. This dual role of separase permits concurrent control of these events for accurate metaphase to anaphase transition. Although the molecular mechanism underlying sister chromatid cohesion has been clarified, that of centriole cohesion is poorly understood. In this study, we show that Akt kinase–interacting protein 1 (Aki1) localizes to centrosomes and regulates centriole cohesion. Aki1 depletion causes formation of multipolar spindles accompanied by centriole splitting, which is separase dependent. We also show that cohesin subunits localize to centrosomes and that centrosomal Scc1 is cleaved by separase coincidentally with chromatin Scc1, suggesting a role of Scc1 as a connector of centrioles as well as sister chromatids. Interestingly, Scc1 depletion strongly induces centriole splitting. Furthermore, Aki1 interacts with cohesin in centrosomes, and this interaction is required for centriole cohesion. We demonstrate that centrosome-associated Aki1 and cohesin play pivotal roles in preventing premature cleavage in centriole cohesion.

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Faculty Opinions recommendation of Structural role of Sfi1p-centrin filaments in budding yeast spindle pole body duplication.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1033701.388954 ◽

2006 ◽

Author(s):

Mark Rose

Keyword(s):

Budding Yeast ◽

Spindle Pole Body ◽

Spindle Pole ◽

Structural Role ◽

Pole Body

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Faculty Opinions recommendation of Fission yeast Myo51 is a meiotic spindle pole body component with discrete roles during cell fusion and spore formation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.2137023.1737135 ◽

2010 ◽

Author(s):

Christopher McInerny

Keyword(s):

Fission Yeast ◽

Cell Fusion ◽

Spindle Pole Body ◽

Spindle Pole ◽

Spore Formation ◽

Meiotic Spindle ◽

Pole Body ◽

Body Component

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Distinct nuclear and spindle pole body populations of cyclin–cdc2 in fission yeast

Nature ◽

10.1038/347680a0 ◽

1990 ◽

Vol 347 (6294) ◽

pp. 680-682 ◽

Cited By ~ 110

Author(s):

Caroline E. Alfa ◽

Bernard Ducommun ◽

David Beach ◽

Jeremy S. Hyams

Keyword(s):

Fission Yeast ◽

Spindle Pole Body ◽

Spindle Pole ◽

Pole Body

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Asymmetry of the spindle pole bodies and spg1p GAP segregation during mitosis in fission yeast

Journal of Cell Science ◽

10.1242/jcs.112.14.2313 ◽

1999 ◽

Vol 112 (14) ◽

pp. 2313-2321 ◽

Cited By ~ 22

Author(s):

L. Cerutti ◽

V. Simanis

Keyword(s):

Cell Cycle ◽

Fission Yeast ◽

Spindle Pole Body ◽

Mitotic Cell ◽

Spindle Pole ◽

The Other ◽

Septum Formation ◽

Spindle Pole Bodies ◽

Interphase Cells ◽

Early Mitosis

In the fission yeast Schizosaccharomyces pombe, the onset of septum formation is induced by a signal transduction network involving several protein kinases and a GTPase switch. One of the roles of the spg1p GTPase is to localise the cdc7p protein kinase to the poles of the mitotic spindle, from where the onset of septation is thought to be signalled at the end of mitosis. Immunofluorescence studies have shown that cdc7p is located on both spindle pole bodies early in mitosis, but only on one during the later stages of anaphase. This is mediated by inactivation of spg1p on one pole before the other. The GAP for spg1p is a complex of two proteins, cdc16p and byr4p. Localisation of cdc16p and byr4p by indirect immunofluorescence during the mitotic cell cycle showed that both proteins are present on the spindle pole body in interphase cells. During mitosis, byr4p is seen first on both poles of the spindle, then on only one. This occurs prior to cdc7p becoming asymmetric. In contrast, the signal due to cdc16p decreases to a low level during early mitosis, before being seen strongly on the same pole as byr4p. Double staining indicates that this is the opposite pole to that which retains cdc7p in late anaphase. Examination of the effect of inactivating cdc16p at various stages of the cell cycle suggests that cdc16p, together with cdc2p plays a role in restraining septum formation during interphase. The asymmetric inactivation of spg1p is mediated by recruitment of the cdc16p-byr4p GAP to one of the poles of the spindle before the other, and the asymmetry of the spindle pole bodies may be established early during mitosis. Moreover, the spindle pole bodies appear to be non-equivalent even after division has been completed.

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The Fission Yeast Homolog of the Human Transcription Factor EAP30 Blocks Meiotic Spindle Pole Body Amplification

Developmental Cell ◽

10.1016/j.devcel.2005.04.016 ◽

2005 ◽

Vol 9 (1) ◽

pp. 63-73 ◽

Cited By ~ 20

Author(s):

Ye Jin ◽

Joel J. Mancuso ◽

Satoru Uzawa ◽

Daniela Cronembold ◽

W. Zacheus Cande

Keyword(s):

Transcription Factor ◽

Fission Yeast ◽

Spindle Pole Body ◽

Spindle Pole ◽

Meiotic Spindle ◽

Pole Body ◽

Yeast Homolog ◽

Human Transcription Factor

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A microtubule polymerase cooperates with the kinesin-6 motor and a microtubule cross-linker to promote bipolar spindle assembly in the absence of kinesin-5 and kinesin-14 in fission yeast

Molecular Biology of the Cell ◽

10.1091/mbc.e17-08-0497 ◽

2017 ◽

Vol 28 (25) ◽

pp. 3647-3659 ◽

Cited By ~ 23

Author(s):

Masashi Yukawa ◽

Tomoki Kawakami ◽

Masaki Okazaki ◽

Kazunori Kume ◽

Ngang Heok Tang ◽

...

Keyword(s):

Fission Yeast ◽

Chromosome Segregation ◽

Spindle Pole Body ◽

Spindle Assembly ◽

Spindle Pole ◽

Temperature Sensitive ◽

Bipolar Spindle ◽

Pole Body ◽

Cross Linker ◽

Spindle Elongation

Accurate chromosome segregation relies on the bipolar mitotic spindle. In many eukaryotes, spindle formation is driven by the plus-end–directed motor kinesin-5 that generates outward force to establish spindle bipolarity. Its inhibition leads to the emergence of monopolar spindles with mitotic arrest. Intriguingly, simultaneous inactivation of the minus-end–directed motor kinesin-14 restores spindle bipolarity in many systems. Here we show that in fission yeast, three independent pathways contribute to spindle bipolarity in the absence of kinesin-5/Cut7 and kinesin-14/Pkl1. One is kinesin-6/Klp9 that engages with spindle elongation once short bipolar spindles assemble. Klp9 also ensures the medial positioning of anaphase spindles to prevent unequal chromosome segregation. Another is the Alp7/TACC-Alp14/TOG microtubule polymerase complex. Temperature-sensitive alp7cut7pkl1 mutants are arrested with either monopolar or very short spindles. Forced targeting of Alp14 to the spindle pole body is sufficient to render alp7cut7pkl1 triply deleted cells viable and promote spindle assembly, indicating that Alp14-mediated microtubule polymerization from the nuclear face of the spindle pole body could generate outward force in place of Cut7 during early mitosis. The third pathway involves the Ase1/PRC1 microtubule cross-linker that stabilizes antiparallel microtubules. Our study, therefore, unveils multifaceted interplay among kinesin-dependent and -independent pathways leading to mitotic bipolar spindle assembly.

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Cytoplasmic microtubular system implicated in de novo formation of a Rabl-like orientation of chromosomes in fission yeast

Journal of Cell Science ◽

10.1242/jcs.114.13.2427 ◽

2001 ◽

Vol 114 (13) ◽

pp. 2427-2435 ◽

Cited By ~ 2

Author(s):

Bunshiro Goto ◽

Koei Okazaki ◽

Osami Niwa

Keyword(s):

Fission Yeast ◽

De Novo ◽

Nuclear Periphery ◽

Spindle Pole Body ◽

Spindle Pole ◽

Limited Area ◽

Cytoplasmic Microtubule ◽

Random Arrangement ◽

Triplex Structure ◽

Microtubular System

Chromosomes are not packed randomly in the nucleus. The Rabl orientation is an example of the non-random arrangement of chromosomes, centromeres are grouped in a limited area near the nuclear periphery and telomeres are located apart from centromeres. This orientation is established during mitosis and maintained through subsequent interphase in a range of species. We report that a Rabl-like configuration can be formed de novo without a preceding mitosis during the transition from the sexual phase to the vegetative phase of the life cycle in fission yeast. In this process, each of the dispersed centromeres is often associated with a novel Sad1-containing body that is contacting a cytoplasmic microtubule laterally (Sad1 is a component of the spindle pole body (SPB)). The Sad1-containing body was colocalized with other known SPB components, Kms1 and Spo15 but not with Cut12, indicating that it represents a novel SPB-related complex. The existence of the triplex structure (centromere-microtubule-Sad1 body) suggests that the clustering of centromeres is controlled by a cytoplasmic microtubular system. Accordingly, when microtubules are destabilized, clustering is markedly reduced.

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Nsk1 ensures accurate chromosome segregation by promoting association of kinetochores to spindle poles during anaphase B

Molecular Biology of the Cell ◽

10.1091/mbc.e11-07-0608 ◽

2011 ◽

Vol 22 (23) ◽

pp. 4486-4502 ◽

Cited By ~ 6

Author(s):

Graham J. Buttrick ◽

John C. Meadows ◽

Theresa C. Lancaster ◽

Vincent Vanoosthuyse ◽

Lindsey A. Shepperd ◽

...

Keyword(s):

Chromosome Segregation ◽

Spindle Pole Body ◽

Spindle Pole ◽

Growth Defect ◽

Catalytic Subunits ◽

Spindle Poles ◽

Sister Chromatids ◽

Anaphase B ◽

Novel Protein

Type 1 phosphatase (PP1) antagonizes Aurora B kinase to stabilize kinetochore–microtubule attachments and to silence the spindle checkpoint. We screened for factors that exacerbate the growth defect of Δdis2 cells, which lack one of two catalytic subunits of PP1 in fission yeast, and identified Nsk1, a novel protein required for accurate chromosome segregation. During interphase, Nsk1 resides in the nucleolus but spreads throughout the nucleoplasm as cells enter mitosis. Following dephosphorylation by Clp1 (Cdc14-like) phosphatase and at least one other phosphatase, Nsk1 localizes to the interface between kinetochores and the inner face of the spindle pole body during anaphase. In the absence of Nsk1, some kinetochores become detached from spindle poles during anaphase B. If this occurs late in anaphase B, then the sister chromatids of unclustered kinetochores segregate to the correct daughter cell. These unclustered kinetochores are efficiently captured, retrieved, bioriented, and segregated during the following mitosis, as long as Dis2 is present. However, if kinetochores are detached from a spindle pole early in anaphase B, then these sister chromatids become missegregated. These data suggest Nsk1 ensures accurate chromosome segregation by promoting the tethering of kinetochores to spindle poles during anaphase B.

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