Sister Kinetochore Recapture in Fission Yeast Occurs by Two Distinct Mechanisms, Both Requiring Dam1 and Klp2

In eukaryotic cells, proper formation of the spindle is necessary for successful cell division. We have studied chromosome recapture in the fission yeast Schizosaccharomyces pombe. We show by live cell analysis that lost kinetochores interact laterally with intranuclear microtubules (INMs) and that both microtubule depolymerization (end-on pulling) and minus-end–directed movement (microtubule sliding) contribute to chromosome retrieval to the spindle pole body (SPB). We find that the minus-end–directed motor Klp2 colocalizes with the kinetochore during its transport to the SPB and contributes to the effectiveness of retrieval by affecting both end-on pulling and lateral sliding. Furthermore, we provide in vivo evidence that Dam1, a component of the DASH complex, also colocalizes with the kinetochore during its transport and is essential for its retrieval by either of these mechanisms. Finally, we find that the position of the unattached kinetochore correlates with the size and orientation of the INMs, suggesting that chromosome recapture may not be a random process.

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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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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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Two-hybrid search for proteins that interact with Sad1 and Kms1, two membrane-bound components of the spindle pole body in fission yeast

Molecular Genetics and Genomics ◽

10.1007/s00438-003-0938-8 ◽

2003 ◽

Vol 270 (6) ◽

pp. 449-461 ◽

Cited By ~ 74

Author(s):

F. Miki ◽

A. Kurabayashi ◽

Y. Tange ◽

K. Okazaki ◽

M. Shimanuki ◽

...

Keyword(s):

Fission Yeast ◽

Spindle Pole Body ◽

Spindle Pole ◽

Hybrid Search ◽

Pole Body ◽

Membrane Bound ◽

Two Hybrid

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Cut1/separase C-terminus affects spindle pole body positioning in interphase of fission yeast: pointed nuclear formation

Genes to Cells ◽

10.1046/j.1365-2443.2002.00586.x ◽

2002 ◽

Vol 7 (11) ◽

pp. 1113-1124 ◽

Cited By ~ 9

Author(s):

Takahiro Nakamura ◽

Koji Nagao ◽

Yukinobu Nakaseko ◽

Mitsuhiro Yanagida

Keyword(s):

Fission Yeast ◽

Spindle Pole Body ◽

Spindle Pole ◽

Pole Body ◽

C Terminus ◽

Body Positioning

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Spindle Pole Body Duplication in Fission Yeast Occurs at the G1/S Boundary but Maturation Is Blocked until Exit from S by an Event Downstream of Cdc10+

Molecular Biology of the Cell ◽

10.1091/mbc.e04-03-0255 ◽

2004 ◽

Vol 15 (12) ◽

pp. 5219-5230 ◽

Cited By ~ 31

Author(s):

Satoru Uzawa ◽

Fei Li ◽

Ye Jin ◽

Kent L. McDonald ◽

Michael B. Braunfeld ◽

...

Keyword(s):

Fission Yeast ◽

Spindle Pole Body ◽

Specific Inhibitor ◽

Spindle Pole ◽

Feedback Mechanism ◽

G1 Arrest ◽

Restrictive Temperature ◽

Permissive Temperature ◽

Pole Body ◽

In Cells

The regulation and timing of spindle pole body (SPB) duplication and maturation in fission yeast was examined by transmission electron microscopy. When cells are arrested at G1 by nitrogen starvation, the SPB is unduplicated. On release from G1, the SPBs were duplicated after 1–2 h. In cells arrested at S by hydroxyurea, SPBs are duplicated but not mature. In G1 arrest/release experiments with cdc2.33 cells at the restrictive temperature, SPBs remained single, whereas in cells at the permissive temperature, SPBs were duplicated. In cdc10 mutant cells, the SPBs seem not only to be duplicated but also to undergo partial maturation, including invagination of the nuclear envelope underneath the SPB. There may be an S-phase–specific inhibitor of SPB maturation whose expression is under control of cdc10+. This model was examined by induction of overreplication of the genome by overexpression of rum1p or cdc18p. In cdc18p-overexpressing cells, the SPBs are duplicated but not mature, suggesting that cdc18p is one component of this feedback mechanism. In contrast, cells overexpressing rum1p have large, deformed SPBs accompanied by other features of maturation and duplication. We propose a feedback mechanism for maturation of the SPB that is coupled with exit from S to trigger morphological changes.

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