scholarly journals Membrane proteins Bqt3 and -4 anchor telomeres to the nuclear envelope to ensure chromosomal bouquet formation

2009 ◽  
Vol 187 (3) ◽  
pp. 413-427 ◽  
Author(s):  
Yuji Chikashige ◽  
Miho Yamane ◽  
Kasumi Okamasa ◽  
Chihiro Tsutsumi ◽  
Tomoko Kojidani ◽  
...  
Keyword(s):  
Nuclear Envelope ◽  
Fission Yeast ◽  
Nuclear Membrane ◽  
Meiotic Prophase ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Null Mutant ◽  
Vegetative Cells ◽  
Nuclear Membranes ◽  
Pole Body

In many organisms, telomeres cluster to form a bouquet arrangement of chromosomes during meiotic prophase. Previously, we reported that two meiotic proteins, Bqt1 and -2, are required for tethering telomeres to the spindle pole body (SPB) during meiotic prophase in fission yeast. This study has further identified two novel, ubiquitously expressed inner nuclear membrane (INM) proteins, Bqt3 and -4, which are required for bouquet formation. We found that in the absence of Bqt4, telomeres failed to associate with the nuclear membranes in vegetative cells and consequently failed to cluster to the SPB in meiotic prophase. In the absence of Bqt3, Bqt4 protein was degraded during meiosis, leading to a phenotype similar to that of the bqt4-null mutant. Collectively, these results show that Bqt4 anchors telomeres to the INM and that Bqt3 protects Bqt4 from protein degradation. Interestingly, the functional integrity of telomeres is maintained even when they are separated from the nuclear envelope in vegetative cells.

scholarly journals LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in fission yeast and human cells

2017 ◽  
Vol 114 (11) ◽  
pp. E2166-E2175 ◽  
Author(s):  
Mingyu Gu ◽  
Dollie LaJoie ◽  
Opal S. Chen ◽  
Alexander von Appen ◽  
Mark S. Ladinsky ◽  
...  
Keyword(s):  
Nuclear Envelope ◽  
Fission Yeast ◽  
Nuclear Membrane ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Human Cells ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Loss Of Function

Endosomal sorting complexes required for transport III (ESCRT-III) proteins have been implicated in sealing the nuclear envelope in mammals, spindle pole body dynamics in fission yeast, and surveillance of defective nuclear pore complexes in budding yeast. Here, we report that Lem2p (LEM2), a member of the LEM (Lap2-Emerin-Man1) family of inner nuclear membrane proteins, and the ESCRT-II/ESCRT-III hybrid protein Cmp7p (CHMP7), work together to recruit additional ESCRT-III proteins to holes in the nuclear membrane. InSchizosaccharomyces pombe, deletion of the ATPasevps4leads to severe defects in nuclear morphology and integrity. These phenotypes are suppressed by loss-of-function mutations that arise spontaneously inlem2orcmp7, implying that these proteins may function upstream in the same pathway. Building on these genetic interactions, we explored the role of LEM2 during nuclear envelope reformation in human cells. We found that CHMP7 and LEM2 enrich at the same region of the chromatin disk periphery during this window of cell division and that CHMP7 can bind directly to the C-terminal domain of LEM2 in vitro. We further found that, during nuclear envelope formation, recruitment of the ESCRT factors CHMP7, CHMP2A, and IST1/CHMP8 all depend on LEM2 in human cells. We conclude that Lem2p/LEM2 is a conserved nuclear site-specific adaptor that recruits Cmp7p/CHMP7 and downstream ESCRT factors to the nuclear envelope.

A calcineurin-like gene ppb1+ in fission yeast: mutant defects in cytokinesis, cell polarity, mating and spindle pole body positioning

1994 ◽  
Vol 107 (7) ◽  
pp. 1725-1735 ◽  
Author(s):  
T. Yoshida ◽  
T. Toda ◽  
M. Yanagida
Keyword(s):  
Cyclosporin A ◽  
Fission Yeast ◽  
Cell Polarity ◽  
Double Mutant ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Predicted Amino Acid Sequence ◽  
Null Mutant ◽  
Pole Body ◽  
Body Positioning

A calcineurin (type 2B)-like protein phosphatase gene designated ppb1+ was isolated from the fission yeast Schizosaccharomyces pombe. The predicted amino acid sequence was 57% identical to rat PP2B alpha. ppb1 null mutant could form colonies at 33 degrees C but the size of the colonies was small at 22 degrees C. Cytokinesis was greatly delayed at 22 degrees C, and a large number of multi-septate cells were produced. The cell polarity control was impaired, causing branched cells. ppb1 null was virtually sterile. These phenotypes were rescued by a plasmid carrying the ppb1+ gene. Multi-septate cells were also produced in wild type at 22 degrees C by cyclosporin A, an inhibitor of calcineurin. This drug effect was enhanced in stst1 null mutant, which was hypersensitive to various drugs and cations. ppb1 null was not affected by cyclosporin A, consistent with the hypothesis that ppb1 is its target. Double-mutant analysis indicated that ppb1 had a function related to that of two other phosphatases, type 1-like dis2 and 2A-like ppa2.ppb1 null-sts1 null showed the severe multi-septate phenotype in the absence of cyclosporin A. ppb1+ and sts1+ gene functions are related. The double mutant ppb1-sts5 was lethal, indicating that the ppb1+ gene shared an essential function with the sts5+ gene. Overexpression of ppb1+ caused anomalies in cell and nuclear shape, microtubule arrays and spindle pole body positioning in interphase cells. Thus the ppb1+ gene appears to be involved in cytokinesis, mating, transport, nuclear and spindle pole body positioning, and cell shape.

scholarly journals Spindle pole body components are reorganized during fission yeast meiosis

2012 ◽  
Vol 23 (10) ◽  
pp. 1799-1811 ◽  
Author(s):  
Midori Ohta ◽  
Masamitsu Sato ◽  
Masayuki Yamamoto
Keyword(s):  
Fission Yeast ◽  
Meiotic Prophase ◽  
Spindle Pole Body ◽  
Mitotic Cell ◽  
Spindle Pole ◽  
Nuclear Movement ◽  
Pole Body ◽  
Proper Number ◽  
Body Components ◽  
Yeast Meiosis

During meiosis, the centrosome/spindle pole body (SPB) must be regulated in a manner distinct from that of mitosis to achieve a specialized cell division that will produce gametes. In this paper, we demonstrate that several SPB components are localized to SPBs in a meiosis-specific manner in the fission yeast Schizosaccharomyces pombe. SPB components, such as Cut12, Pcp1, and Spo15, which stay on the SPB during the mitotic cell cycle, disassociate from the SPB during meiotic prophase and then return to the SPB immediately before the onset of meiosis I. Interestingly, the polo kinase Plo1, which normally localizes to the SPB during mitosis, is excluded from them in meiotic prophase, when meiosis-specific, horse-tail nuclear movement occurs. We found that exclusion of Plo1 during this period was essential to properly remodel SPBs, because artificial targeting of Plo1 to SPBs resulted in an overduplication of SPBs. We also found that the centrin Cdc31 was required for meiotic SPB remodeling. Thus Plo1 and a centrin play central roles in the meiotic SPB remodeling, which is essential for generating the proper number of meiotic SPBs and, thereby provide unique characteristics to meiotic divisions.

scholarly journals The SUN Protein Mps3 Is Required for Spindle Pole Body Insertion into the Nuclear Membrane and Nuclear Envelope Homeostasis

PLoS Genetics ◽  
2011 ◽  
Vol 7 (11) ◽  
pp. e1002365 ◽  
Author(s):  
Jennifer M. Friederichs ◽  
Suman Ghosh ◽  
Christine J. Smoyer ◽  
Scott McCroskey ◽  
Brandon D. Miller ◽  
...  
Keyword(s):  
Nuclear Envelope ◽  
Nuclear Membrane ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
The Sun ◽  
Pole Body

scholarly journals Dissociation of the Nuf2-Ndc80 Complex Releases Centromeres from the Spindle-Pole Body during Meiotic Prophase in Fission Yeast

2005 ◽  
Vol 16 (5) ◽  
pp. 2325-2338 ◽  
Author(s):  
Haruhiko Asakawa ◽  
Aki Hayashi ◽  
Tokuko Haraguchi ◽  
Yasushi Hiraoka
Keyword(s):  
Fission Yeast ◽  
Chromosome Segregation ◽  
Meiotic Prophase ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Mating Pheromone ◽  
Pole Body ◽  
Pheromone Signaling ◽  
Ndc80 Complex ◽  
Mutant Cells

In the fission yeast Schizosaccharomyces pombe, centromeres remain clustered at the spindle-pole body (SPB) during mitotic interphase. In contrast, during meiotic prophase centromeres dissociate from the SPB. Here we examined the behavior of centromere proteins in living meiotic cells of S. pombe. We show that the Nuf2-Ndc80 complex proteins (Nuf2, Ndc80, Spc24, and Spc25) disappear from the centromere in meiotic prophase when the centromeres are separated from the SPB. The centromere protein Mis12 also dissociates during meiotic prophase; however, Mis6 remains throughout meiosis. When cells are induced to meiosis by inactivation of Pat1 kinase (a key negative regulator of meiosis), centromeres remain associated with the SPB during meiotic prophase. However, inactivation of Nuf2 by a mutation causes the release of centromeres from the SPB in pat1 mutant cells, suggesting that the Nuf2-Ndc80 complex connects centromeres to the SPB. We further found that removal of the Nuf2-Ndc80 complex from the centromere and centromere-SPB dissociation are caused by mating pheromone signaling. Because pat1 mutant cells also show aberrant chromosome segregation in the first meiotic division and this aberration is compensated by mating pheromone signaling, dissociation of the Nuf2-Ndc80 complex may be associated with remodeling of the kinetochore for meiotic chromosome segregation.

scholarly journals Bqt2p is essential for initiating telomere clustering upon pheromone sensing in fission yeast

2006 ◽  
Vol 173 (6) ◽  
pp. 845-851 ◽  
Author(s):  
Xie Tang ◽  
Ye Jin ◽  
W. Zacheus Cande
Keyword(s):  
Fission Yeast ◽  
Chromosome Pairing ◽  
Meiotic Prophase ◽  
Homologous Chromosome ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Meiotic Spindle ◽  
Meiotic Progression ◽  
Pole Body ◽  
Homologous Chromosome Pairing

The telomere bouquet, i.e., telomere clustering on the nuclear envelope (NE) during meiotic prophase, is thought to promote homologous chromosome pairing. Using a visual screen, we identified bqt2/im295, a mutant that disrupts telomere clustering in fission yeast. Bqt2p is required for linking telomeres to the meiotic spindle pole body (SPB) but not for attachment of telomeres or the SPB to the NE. Bqt2p is expressed upon pheromone sensing and colocalizes thereafter to Sad1p, an SPB protein. This localization only depends on Bqt1p, not on other identified proteins required for telomere clustering. Upon pheromone sensing, generation of Sad1p foci next to telomeres depends on Bqt2p. However, depletion of Bqt2p from the SPB is dispensable for dissolving the telomere bouquet at the end of meiotic prophase. Therefore, telomere bouquet formation requires Bqt2p as a linking component and is finely regulated during meiotic progression.

scholarly journals The product of the spindle formation gene sad1+ associates with the fission yeast spindle pole body and is essential for viability.

1995 ◽  
Vol 129 (4) ◽  
pp. 1033-1047 ◽  
Author(s):  
I Hagan ◽  
M Yanagida
Keyword(s):  
Fission Yeast ◽  
Nuclear Membrane ◽  
Essential Gene ◽  
Nuclear Periphery ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Temperature Sensitive ◽  
Spindle Formation ◽  
Isolation And Characterization ◽  
Pole Body

Spindle formation in fission yeast occurs by the interdigitation of two microtubule arrays extending from duplicated spindle pole bodies which span the nuclear membrane. By screening a bank of temperature-sensitive mutants by anti-tubulin immunofluorescence microscopy, we previously identified the sad1.1 mutation (Hagan, I., and M. Yanagida. 1990. Nature (Lond.). 347:563-566). Here we describe the isolation and characterization of the sad1+ gene. We show that the sad1.1 mutation affected both spindle formation and function. The sad1+ gene is a novel essential gene that encodes a protein with a predicted molecular mass of 58 kD. Deletion of the gene was lethal resulting in identical phenotypes to the sad1.1 mutation. Sequence analysis predicted a potential membrane-spanning domain and an acidic amino terminus. Sad1 protein migrated as two bands of 82 and 84 kD on SDS-PAGE, considerably slower than its predicted mobility, and was exclusively associated with the spindle pole body (SPB) throughout the mitotic and meiotic cycles. Microtubule integrity was not required for Sad1 association with the SPB. Upon the differentiation of the SPB in metaphase of meiosis II, Sad1-staining patterns similarly changed from a dot to a crescent supporting an integral role in SPB function. Moderate overexpression of Sad1 led to association with the nuclear periphery. As Sad1 was not detected in the cytoplasmic microtubule-organizing centers activated at the end of anaphase or kinetochores, we suggest that Sad1 is not a general component of microtubule-interacting structures per se, but is an essential mitotic component that associates with the SPB but is not required for microtubule nucleation. Sad1 may play a role in SPB structure, such as maintaining a functional interface with the nuclear membrane or in providing an anchor for the attachment of microtubule motor proteins.

scholarly journals LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in fission yeast and human cells

2016 ◽  
Author(s):  
Mingyu Gu ◽  
Dollie LaJoie ◽  
Opal S. Chen ◽  
Alexander Von Appen ◽  
Mark S. Ladinsky ◽  
...  
Keyword(s):  
Nuclear Envelope ◽  
Fission Yeast ◽  
Nuclear Membrane ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Human Cells ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Loss Of Function

AbstractESCRT-III proteins have been implicated in sealing the nuclear envelope in mammals, spindle pole body dynamics in fission yeast, and surveillance of defective nuclear pore complexes in budding yeast. Here, we report that Lem2p (LEM2), a member of the LEM (Lap2-Emerin-Man1) family of inner nuclear membrane proteins, and the ESCRT-II/ESCRT-III hybrid protein Cmp7p (CHMP7), work together to recruit additional ESCRT-III proteins to holes in the nuclear membrane. InS. pombe, deletion of the ATPasevps4leads to severe defects in nuclear morphology and integrity. These phenotypes are suppressed by loss-of-function mutations that arise spontaneously inlem2orcmp7,implying that these proteins may function upstream in the same pathway. Building on these genetic interactions, we explored the role of LEM2 during nuclear envelope reformation in human cells. We found that CHMP7 and LEM2 enrich at the same region of the chromatin disc periphery during this window of cell division, and that CHMP7 can bind directly to the C-terminal domain of LEM2in vitro. We further found that, during nuclear envelope formation, recruitment of the ESCRT factors CHMP7, CHMP2A and IST1/CHMP8 all depend on LEM2 in human cells. We conclude that Lem2p/LEM2 is a conserved nuclear site-specific adaptor that recruits Cmp7p/CHMP7 and downstream ESCRT factors to the nuclear envelope.

Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells

1998 ◽  
Vol 111 (6) ◽  
pp. 701-712 ◽  
Author(s):  
D.Q. Ding ◽  
Y. Chikashige ◽  
T. Haraguchi ◽  
Y. Hiraoka
Keyword(s):  
Fission Yeast ◽  
Meiotic Prophase ◽  
Fluorescent Protein ◽  
Dynamic Instability ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Living Cells ◽  
Continuous Observation ◽  
Nuclear Movement ◽  
Pole Body

Using a computerized fluorescence microscope system to observe fluorescently stained cellular structures in vivo, we have examined the dynamics of chromosomes and microtubules during the process of meiosis in the fission yeast Schizosaccharomyces pombe. Fission yeast meiotic prophase is characterized by a distinctive type of nuclear movement that is led by telomeres clustered at the spindle-pole body (the centrosome-equivalent structure in fungi): the nucleus oscillates back and forth along the cell axis, moving continuously between the two ends of the cell for some hours prior to the meiotic divisions. To obtain a dynamic view of this oscillatory nuclear movement in meiotic prophase, we visualized microtubules and chromosomes in living cells using jellyfish green fluorescent protein fused with alpha-tubulin and a DNA-specific fluorescent dye, Hoechst 33342, respectively. Continuous observation of chromosomes and microtubules in these cells demonstrated that the oscillatory nuclear movement is mediated by dynamic reorganization of astral microtubules originating from the spindle-pole body. During each half-oscillatory period, the microtubules extending rearward from the leading edge of the nucleus elongate to drive the nucleus to one end of the cell. When the nucleus reversed direction, its motion during the second half of the oscillation was not driven by the same microtubules that drove its motion during the first half, but rather by newly assembled microtubules. Reversible inhibition of nuclear movement by an inhibitor of microtubule polymerization, thiabendazole, confirmed the involvement of astral microtubules in oscillatory nuclear movement. The speed of the movement fluctuated within a range 0 to 15 micron/minute, with an average of about 5 microm/minute. We propose a model in which the oscillatory nuclear movement is mediated by dynamic instability and selective stabilization of astral microtubules.

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