Mps1 promotes poleward chromosome movements in meiotic pro-metaphase

Data Support ◽

Chromosome Movements ◽

Meiosis I ◽

Prophase Of Meiosis

ABSTRACTIn prophase of meiosis I, homologous partner chromosomes pair and become connected by crossovers. Chiasmata, the connections formed between the partners enable the chromosome pair, called a bivalent, to attach as a single unit to the spindle. When the meiosis I spindle forms in prometaphase, most bivalents are associated with a single spindle pole and go through a series of oscillations on the spindle, attaching to and detaching from microtubules until the partners of the bivalent are bi-oriented, that is, attached to microtubules from opposite sides of the spindle, and prepared to be segregated at anaphase I. The conserved, kinetochore-associated kinase, Mps1, is essential for the bivalents to be pulled by microtubules across the spindle in prometaphase. Here we show that MPS1 is not required for kinetochores to attach microtubules but instead is necessary to trigger the migration of microtubule-attached kinetochores towards the poles. Our data support the model that Mps1 triggers depolymerization of microtubule ends once they attach to kinetochores in prometaphase. Thus, Mps1 acts at the kinetochore to co-ordinate the successful attachment of a microtubule and the triggering of microtubule depolymerization to move the chromosome.

Mps1 promotes poleward chromosome movements in meiotic prometaphase

Molecular Biology of the Cell ◽

10.1091/mbc.e20-08-0525-t ◽

2021 ◽

Vol 32 (10) ◽

pp. 1020-1032

Author(s):

Régis E. Meyer ◽

Aaron R. Tipton ◽

Rebecca LaVictoire ◽

Gary J. Gorbsky ◽

Dean S. Dawson

Keyword(s):

Spindle Checkpoint ◽

Spindle Pole ◽

Microtubule Attachment ◽

Mitosis And Meiosis ◽

Chromosome Movements

Mps1 is a kinase that regulates several steps in mitosis and meiosis. Mps1 is essential for the spindle checkpoint and helps stabilize attachment of kinetochores to microtubules. Here we show that following microtubule attachment, Mps1 promotes microtubule depolymerization to trigger migration of the chromosome toward the spindle pole.

Paired arrangement of kinetochores together with microtubule pivoting and dynamics drive kinetochore capture in meiosis I

Scientific Reports ◽

10.1038/srep25736 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 10

Author(s):

Gheorghe Cojoc ◽

Ana-Maria Florescu ◽

Alexander Krull ◽

Anna H. Klemm ◽

Nenad Pavin ◽

...

Keyword(s):

Cell Division ◽

Fission Yeast ◽

General Feature ◽

Protein Complexes ◽

Spindle Pole ◽

Single Object ◽

Homologous Chromosomes ◽

Angular Movement ◽

Spindle Microtubules ◽

Meiosis I

Abstract Kinetochores are protein complexes on the chromosomes, whose function as linkers between spindle microtubules and chromosomes is crucial for proper cell division. The mechanisms that facilitate kinetochore capture by microtubules are still unclear. In the present study, we combine experiments and theory to explore the mechanisms of kinetochore capture at the onset of meiosis I in fission yeast. We show that kinetochores on homologous chromosomes move together, microtubules are dynamic and pivot around the spindle pole, and the average capture time is 3–4 minutes. Our theory describes paired kinetochores on homologous chromosomes as a single object, as well as angular movement of microtubules and their dynamics. For the experimentally measured parameters, the model reproduces the measured capture kinetics and shows that the paired configuration of kinetochores accelerates capture, whereas microtubule pivoting and dynamics have a smaller contribution. Kinetochore pairing may be a general feature that increases capture efficiency in meiotic cells.

A permeabilized cell model for studying cell division: a comparison of anaphase chromosome movement and cleavage furrow constriction in lysed PtK1 cells.

10.1083/jcb.88.3.618 ◽

1981 ◽

Vol 88 (3) ◽

pp. 618-629 ◽

Cited By ~ 50

Author(s):

W Z Cande ◽

K McDonald ◽

R L Meeusen

Keyword(s):

Cell Model ◽

Chromosome Movement ◽

Mitotic Apparatus ◽

Permeabilized Cell ◽

Culture Cells ◽

Brij 58 ◽

Myosin Subfragment ◽

Myosin Subfragment 1 ◽

Chromosome Movements

After lysis in a Brij 58-polyethylene glycol medium, PtK1 cells are permeable to small molecules, such as erythrosin B, and to proteins, such as rhodamine-labeled FAB, myosin subfragment-1, and tubulin. Holes are present in the plasma membrane, and the mitochondria are swollen and distorted, but other membrane-bounded organelles of the lysed cell model are not noticeably altered. After lysis, the mitotic apparatus is functional; chromosomes move poleward and the spindle elongates. Cells lysed while in cytokinesis will continue to divide for several minutes. Addition of crude tubulin extracts, MAP-free tubulin, or taxol to the lysis medium retards anaphase chromosome movements but does not affect cleavage. On the other hand, N-ethylmaleimide-modified myosin subfragment-1, phalloidin, and cytochalasin B inhibit cleavage but have no effect on anaphase chromosome movements under identical lysis conditions. These results suggest that actomyosin plays no functional role in anaphase chromosome movement in mammalian tissue culture cells and that microtubule depolymerization is a rate-limiting step for chromosome-to-pole movements.

DIPLOID SPORE FORMATION AND OTHER MEIOTIC EFFECTS OF TWO CELL-DIVISION-CYCLE MUTATIONS OF SACCHAROMYCES CEREVISIAE

Genetics ◽

10.1093/genetics/96.4.859 ◽

1980 ◽

Vol 96 (4) ◽

pp. 859-876 ◽

Cited By ~ 3

Author(s):

David Schild ◽

Breck Byers

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Division ◽

Spindle Pole Body ◽

Spindle Pole ◽

Cell Division Cycle ◽

Temperature Sensitive ◽

Restrictive Temperature ◽

Single Spore ◽

Diploid Cells ◽

Meiosis I

ABSTRACT The meiotic effects of two cell-division-cycle mutations of Saccharomyces cerevisiae (cdc5 and cdc14) have been examined. These mutations were isolated by L. H. Hartwell and his colleagues and characterized as defective in mitosis, causing a temperature-sensitive arrest in late nuclear division. When subjected to the restrictive temperature in meiosis, diploid cells homozygous for either of these mutations generally proceeded through premeiotic DNA synthesis and commitment to meiotic levels of recombination, but then arrested at a stage following spindle pole body (SPB) duplication and separation. The two SPBs lacked the interconnection by spindle microtubules typical of the complete meiosis I spindle. Challenge of these homozygotes by a semi-restrictive temperature often caused the production of asci containing two diploid spores. Genetic analysis of the viable pairs of spores revealed that each spore had become homozygous for centromere-linked markers significantly more frequently than for distal markers, indicating that the two spores each contained pairs of sister centromeres that had co-segregated in the reductional division of meiosis I. Ultrastructural analysis of the cdc5 homozygote demonstrated that these cells had completed meiosis I and formed two meiosis II spindles, but that the latter remained unusually short. This resulted in the encapsulation of both poles of each spindle within a single spore wall. These mutations therefore are defective in both meiotic divisions, as well as in the mitotic division described originally.

Kinetochore-mediated outward force promotes spindle pole separation in fission yeast

Molecular Biology of the Cell ◽

10.1091/mbc.e19-07-0366 ◽

2019 ◽

Vol 30 (22) ◽

pp. 2802-2813 ◽

Cited By ~ 4

Author(s):

Yutaka Shirasugi ◽

Masamitsu Sato

Keyword(s):

Fission Yeast ◽

Motor Proteins ◽

Spindle Assembly ◽

Spindle Pole ◽

Double Knockout ◽

Bipolar Spindle ◽

Spindle Poles ◽

Bipolar Spindles ◽

Meiosis I

Bipolar spindles are organized by motor proteins that generate microtubule-dependent forces to separate the two spindle poles. The fission yeast Cut7 (kinesin-5) is a plus-end-directed motor that generates the outward force to separate the two spindle poles, whereas the minus-end-directed motor Pkl1 (kinesin-14) generates the inward force. Balanced forces by these antagonizing kinesins are essential for bipolar spindle organization in mitosis. Here, we demonstrate that chromosomes generate another outward force that contributes to the bipolar spindle assembly. First, it was noted that the cut7 pkl1 double knockout failed to separate spindle poles in meiosis I, although the mutant is known to succeed it in mitosis. It was assumed that this might be because meiotic kinetochores of bivalent chromosomes joined by cross-overs generate weaker tensions in meiosis I than the strong tensions in mitosis generated by tightly tethered sister kinetochores. In line with this idea, when meiotic mono-oriented kinetochores were artificially converted to a mitotic bioriented layout, the cut7 pkl1 mutant successfully separated spindle poles in meiosis I. Therefore, we propose that spindle pole separation is promoted by outward forces transmitted from kinetochores to spindle poles through microtubules.

The Saccharomyces cerevisiae Kinesin-related Motor Kar3p Acts at Preanaphase Spindle Poles to Limit the Number and Length of Cytoplasmic Microtubules

10.1083/jcb.137.2.417 ◽

1997 ◽

Vol 137 (2) ◽

pp. 417-431 ◽

Cited By ~ 89

Author(s):

William Saunders ◽

David Hornack ◽

Valerie Lengyel ◽

Changchun Deng

Keyword(s):

Saccharomyces Cerevisiae ◽

Spindle Pole Body ◽

Spindle Pole ◽

Body Region ◽

Growth Defects ◽

Microtubule Polymerization ◽

Late Anaphase ◽

A Cell ◽

Cytoplasmic Microtubules

The Saccharomyces cerevisiae kinesin-related motor Kar3p, though known to be required for karyogamy, plays a poorly defined, nonessential role during vegetative growth. We have found evidence suggesting that Kar3p functions to limit the number and length of cytoplasmic microtubules in a cell cycle–specific manner. Deletion of KAR3 leads to a dramatic increase in cytoplasmic microtubules, a phenotype which is most pronounced from START through the onset of anaphase but less so during late anaphase in synchronized cultures. We have immunolocalized HA-tagged Kar3p to the spindle pole body region, and fittingly, Kar3p was not detected by late anaphase. A microtubule depolymerizing activity may be the major vegetative role for Kar3p. Addition of the microtubule polymerization inhibitors nocodazol or benomyl to the medium or deletion of the nonessential α-tubulin TUB3 gene can mostly correct the abnormal microtubule arrays and other growth defects of kar3 mutants, suggesting that these phenotypes result from excessive microtubule polymerization. Microtubule depolymerization may also be the mechanism by which Kar3p acts in opposition to the anaphase B motors Cin8p and Kip1p. A preanaphase spindle collapse phenotype of cin8 kip1 mutants, previously shown to involve Kar3p, is markedly delayed when microtubule depolymerization is inhibited by the tub2-150 mutation. These results suggest that the Kar3p motor may act to regulate the length and number of microtubules in the preanaphase spindle.

Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells.

10.1083/jcb.110.1.81 ◽

1990 ◽

Vol 110 (1) ◽

pp. 81-95 ◽

Cited By ~ 291

Author(s):

C L Rieder ◽

S P Alexander

Keyword(s):

Spatial Separation ◽

Time Lapse ◽

Spindle Pole ◽

Variable Number ◽

Fiber Formation ◽

Polar Regions ◽

Chromosome Attachment ◽

Kinetochore Region ◽

Tubulin Immunofluorescence

During mitosis in cultured newt pneumocytes, one or more chromosomes may become positioned well removed (greater than 50 microns) from the polar regions during early prometaphase. As a result, these chromosomes are delayed for up to 5 h in forming an attachment to the spindle. The spatial separation of these chromosomes from the polar microtubule-nucleating centers provides a unique opportunity to study the initial stages of kinetochore fiber formation in living cells. Time-lapse Nomarski-differential interference contrast videomicroscopic observations reveal that late-attaching chromosomes always move, upon attachment, into a single polar region (usually the one closest to the chromosome). During this attachment, the kinetochore region of the chromosome undergoes a variable number of transient poleward tugs that are followed, shortly thereafter, by rapid movement of the chromosome towards the pole. Anti-tubulin immunofluorescence and serial section EM reveal that the kinetochores and kinetochore regions of nonattached chromosomes lack associated microtubules. By contrast, these methods reveal that the attachment and subsequent poleward movement of a chromosome correlates with the association of a single long microtubule with one of the kinetochores of the chromosome. This microtubule traverses the entire distance between the spindle pole and the kinetochore and often extends well past the kinetochore. From these results, we conclude that the initial attachment of a chromosome to the newt pneumocyte spindle results from an interaction between a single polar-nucleated microtubule and one of the kinetochores on the chromosome. Once this association is established, the kinetochore is rapidly transported poleward along the surface of the microtubule by a mechanism that is not dependent on microtubule depolymerization. Our results further demonstrate that the motors for prometaphase chromosome movement must be either on the surface of the kinetochore (i.e., within the corona but not the plate), distributed along the surface of the kinetochore microtubules, or both.

The ultrastructure of nuclear division in Armillaria mellea: meiosis

Canadian Journal of Botany ◽

10.1139/b79-235 ◽

1979 ◽

Vol 57 (18) ◽

pp. 1860-1872 ◽

Cited By ~ 12

Author(s):

Diane Cope Peabody ◽

Jerome J. Motta

Keyword(s):

Nuclear Envelope ◽

Membrane Structure ◽

Nuclear Division ◽

Spindle Pole ◽

Armillaria Mellea ◽

Spindle Pole Bodies ◽

Late Telophase ◽

Meiosis I ◽

Metaphase I

Meiosis I in isolates of Armillaria mellea in which subhymenial hyphae are uninucleate and lack clamp connections was examined ultrastructurally. Although the overall pattern of development and basidiosporogenesis appears similar to other Homobasidiomycetes it was observed that spindle pole bodies are predominantly monoglobular and are associated with a unique membrane structure of the subtending nuclear envelope. The nuclear envelope also disappears at metaphase I and reforms by the coalescence of membrane fragments around the compacted chromatin at late telophase I. The significance of these features in relation to other Basidiomycetes is briefly discussed.

The Drosophila Kinesin-13, KLP59D, Impacts Pacman- and Flux-based Chromosome Movement

Molecular Biology of the Cell ◽

10.1091/mbc.e09-07-0557 ◽

2009 ◽

Vol 20 (22) ◽

pp. 4696-4705 ◽

Cited By ~ 20

Author(s):

Uttama Rath ◽

Gregory C. Rogers ◽

Dongyan Tan ◽

Maria Ana Gomez-Ferreria ◽

Daniel W. Buster ◽

...

Keyword(s):

Electron Microscopy ◽

Family Member ◽

Somatic Cells ◽

Spindle Pole ◽

The Other ◽

Chromosome Movement ◽

Spindle Microtubule ◽

Important Regulator ◽

Chromosome Movements

Chromosome movements are linked to the active depolymerization of spindle microtubule (MT) ends. Here we identify the kinesin-13 family member, KLP59D, as a novel and uniquely important regulator of spindle MT dynamics and chromosome motility in Drosophila somatic cells. During prometaphase and metaphase, depletion of KLP59D, which targets to centrosomes and outer kinetochores, suppresses the depolymerization of spindle pole–associated MT minus ends, thereby inhibiting poleward tubulin Flux. Subsequently, during anaphase, loss of KLP59D strongly attenuates chromatid-to-pole motion by suppressing the depolymerization of both minus and plus ends of kinetochore-associated MTs. The mechanism of KLP59D's impact on spindle MT plus and minus ends appears to differ. Our data support a model in which KLP59D directly depolymerizes kinetochore-associated plus ends during anaphase, but influences minus ends indirectly by localizing the pole-associated MT depolymerase KLP10A. Finally, electron microscopy indicates that, unlike the other Drosophila kinesin-13s, KLP59D is largely incapable of oligomerizing into MT-associated rings in vitro, suggesting that such structures are not a requisite feature of kinetochore-based MT disassembly and chromosome movements.

Cell division in two large pennate diatoms Hantzschia and Nitzschia III. A new proposal for kinetochore function during prometaphase.

10.1083/jcb.86.2.402 ◽

1980 ◽

Vol 86 (2) ◽

pp. 402-416 ◽

Cited By ~ 58

Author(s):

D H Tippit ◽

J D Pickett-Heaps ◽

R Leslie

Keyword(s):

Electron Microscopy ◽

Leading Edge ◽

Time Lapse ◽

Cell Types ◽

Spindle Pole ◽

Large Species ◽

Conventional View ◽

Chromosome Movements ◽

Kinetochore Function

Prometaphase in two large species of diatoms is examined, using the following techniques: (a) time-lapse cinematography of chromosome movements in vivo; (b) electron microscopy of corresponding stages: (c) reconstruction of the microtubules (MTs) in the kinetochore fiber of chromosomes attached to the spindle. In vivo, the chromosomes independently commence oscillations back and forth to one pole. The kinetochore is usually at the leading edge of such chromosome movements; a variable time later both kinetochores undergo such oscillations but toward opposite poles and soon stretch poleward to establish stable bipolar attachment. Electron microscopy of early prometaphase shows that the kinetochores usually laterally associate with MTs that have one end attached to the spindle pole. At late prometaphase, most chromosomes are fully attached to the spindle, but the kinetochores on unattached chromosomes are bare of MTs. Reconstruction of the kinetochore fiber demonstrates that most of its MTs (96%) extend past the kinetochore and are thus apparently not nucleated there. At least one MT terminates at each kinetochore analyzed. Our interpretation is that the conventional view of kinetochore function cannot apply to diatoms. The kinetochore fiber in diatoms appears to be primarily composed of MTs from the poles, in contrast to the conventional view that many MTs of the kinetochore fiber are nucleated by the kinetochore. Similarly, chromosomes appear to initially orient their kinetochores to opposite poles by moving along MTs attached to the poles, instead of orientation effected by kinetochore MTs laterally associating with other MTs in the spindle. The function of the kinetochore in diatoms and other cell types is discussed.