scholarly journals Regulation of mitosis by poly(ADP-ribosyl)ation

2005 ◽  
Vol 391 (2) ◽  
Author(s):  
Duane A. Compton
Keyword(s):  
Cell Division ◽  
Mammalian Cells ◽  
Protein A ◽  
Mitotic Apparatus ◽  
Spindle Poles ◽  
Biochemical Journal ◽  
Dividing Cells ◽  
Cross Links ◽  
Tankyrase 1 ◽  
The Impact

The spindle is a dynamic, microtubule-based structure responsible for chromosome segregation during cell division. Spindles in mammalian cells contain several thousand microtubules that are arranged into highly symmetric bipolar arrays by the actions of numerous microtubule-associated motor and non-motor proteins. In addition to these protein constituents, recent work has demonstrated that poly(ADP-ribose) is a key spindle component. Of the multitude of poly(ADP-ribose) polymerase proteins encoded in the genome, tankyrase 1 appears to be the primary enzyme responsible for building poly(ADP-ribose) in spindles during mitosis. In this issue of the Biochemical Journal, Susan Smith and co-workers show that the primary target of tankyrase 1 in dividing cells is NuMA (nuclear mitotic apparatus protein), a protein that cross-links microtubule ends at spindle poles. The impact of poly(ADP-ribosyl)ation on the biochemical function of NuMA remains murky at this time, but these new results represent the first step to clearing the view as to how poly(ADP-ribosyl)ation regulates cell division.

scholarly journals Cytokinesis and Midzone Microtubule Organization inCaenorhabditis elegans Require the Kinesin-like Protein ZEN-4

1998 ◽  
Vol 9 (8) ◽  
pp. 2037-2049 ◽  
Author(s):  
William B. Raich ◽  
Adrienne N. Moran ◽  
Joel H. Rothman ◽  
Jeff Hardin
Keyword(s):  
Cell Division ◽  
Null Allele ◽  
Time Lapse ◽  
Equatorial Region ◽  
Microtubule Organization ◽  
Dividing Cells ◽  
Spindle Midzone ◽  
Cross Links ◽  
Complete Cell

Members of the MKLP1 subfamily of kinesin motor proteins localize to the equatorial region of the spindle midzone and are capable of bundling antiparallel microtubules in vitro. Despite these intriguing characteristics, it is unclear what role these kinesins play in dividing cells, particularly within the context of a developing embryo. Here, we report the identification of a null allele ofzen-4, an MKLP1 homologue in the nematodeCaenorhabditis elegans, and demonstrate that ZEN-4 is essential for cytokinesis. Embryos deprived of ZEN-4 form multinucleate single-celled embryos as they continue to cycle through mitosis but fail to complete cell division. Initiation of the cytokinetic furrow occurs at the normal time and place, but furrow propagation halts prematurely. Time-lapse recordings and microtubule staining reveal that the cytokinesis defect is preceded by the dissociation of the midzone microtubules. We show that ZEN-4 protein localizes to the spindle midzone during anaphase and persists at the midbody region throughout cytokinesis. We propose that ZEN-4 directly cross-links the midzone microtubules and suggest that these microtubules are required for the completion of cytokinesis.

Asymmetric cell division in fucoid algae: a role for cortical adhesions in alignment of the mitotic apparatus

2001 ◽  
Vol 114 (23) ◽  
pp. 4319-4328
Author(s):  
Sherryl R. Bisgrove ◽  
Darryl L. Kropf
Keyword(s):  
Cell Division ◽  
Asymmetric Cell Division ◽  
Cell Cortex ◽  
Mitotic Apparatus ◽  
Pharmacological Agents ◽  
Division Plane ◽  
Adhesion Sites ◽  
Spindle Poles ◽  
Pelvetia Compressa ◽  
Two Phases

The first cell division in zygotes of the fucoid brown alga Pelvetia compressa is asymmetric and we are interested in the mechanism controlling the alignment of this division. Since the division plane bisects the mitotic apparatus, we investigated the timing and mechanism of spindle alignments. Centrosomes, which give rise to spindle poles, aligned with the growth axis in two phases – a premetaphase rotation of the nucleus and centrosomes followed by a postmetaphase alignment that coincided with the separation of the mitotic spindle poles during anaphase and telophase. The roles of the cytoskeleton and cell cortex in the two phases of alignment were analyzed by treatment with pharmacological agents. Treatments that disrupted cytoskeleton or perturbed cortical adhesions inhibited pre-metaphase alignment and we propose that this rotational alignment is effected by microtubules anchored at cortical adhesion sites. Postmetaphase alignment was not affected by any of the treatments tested, and may be dependent on asymmetric cell morphology.

scholarly journals Evidence for dynein and astral microtubule–mediated cortical release and transport of Gαi/LGN/NuMA complex in mitotic cells

2013 ◽  
Vol 24 (7) ◽  
pp. 901-913 ◽  
Author(s):  
Zhen Zheng ◽  
Qingwen Wan ◽  
Jing Liu ◽  
Huabin Zhu ◽  
Xiaogang Chu ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Fluorescence Recovery After Photobleaching ◽  
Cytoplasmic Dynein ◽  
Cell Cortex ◽  
Mitotic Apparatus ◽  
Spindle Positioning ◽  
Spindle Poles ◽  
Astral Microtubule ◽  
Mammalian Homologue ◽  
Cortical Localization

Spindle positioning is believed to be governed by the interaction between astral microtubules and the cell cortex and involve cortically anchored motor protein dynein. How dynein is recruited to and regulated at the cell cortex to generate forces on astral microtubules is not clear. Here we show that mammalian homologue of Drosophila Pins (Partner of Inscuteable) (LGN), a Gαi-binding protein that is critical for spindle positioning in different systems, associates with cytoplasmic dynein heavy chain (DYNC1H1) in a Gαi-regulated manner. LGN is required for the mitotic cortical localization of DYNC1H1, which, in turn, also modulates the cortical accumulation of LGN. Using fluorescence recovery after photobleaching analysis, we show that cortical LGN is dynamic and the turnover of LGN relies, at least partially, on astral microtubules and DYNC1H1. We provide evidence for dynein- and astral microtubule–mediated transport of Gαi/LGN/nuclear mitotic apparatus (NuMA) complex from cell cortex to spindle poles and show that actin filaments counteract such transport by maintaining Gαi/LGN/NuMA and dynein at the cell cortex. Our results indicate that astral microtubules are required for establishing bipolar, symmetrical cortical LGN distribution during metaphase. We propose that regulated cortical release and transport of LGN complex along astral microtubules may contribute to spindle positioning in mammalian cells.

scholarly journals Roles of kinesin and kinesin-like proteins in sea urchin embryonic cell division: evaluation using antibody microinjection.

1993 ◽  
Vol 123 (3) ◽  
pp. 681-689 ◽  
Author(s):  
B D Wright ◽  
M Terasaki ◽  
J M Scholey
Keyword(s):  
Cell Division ◽  
Sea Urchin ◽  
Heavy Chain ◽  
Mammalian Cells ◽  
Embryonic Cell ◽  
Mitotic Apparatus ◽  
Specific Antibodies ◽  
Sea Urchin Embryos ◽  
Mitotic Figures

Previous studies suggest that kinesin heavy chain (KHC) is associated with ER-derived membranes that accumulate in the mitotic apparatus in cells of early sea urchin embryos (Wright, B. D., J. H. Henson, K. P. Wedaman, P. J. Willy, J. N. Morand, and J. M. Scholey. 1991. J. Cell Biol. 113:817-833). Here, we report that the microinjection of KHC-specific antibodies into these cells has no effect on mitosis or ER membrane organization, even though one such antibody, SUK4, blocks kinesin-driven motility in vitro and in mammalian cells. Microinjected SUK4 was localized to early mitotic figures, suggesting that it is able to access kinesin in spindles. In contrast to KHC-specific antibodies, two antibodies that react with kinesin-like proteins (KLPs), namely CHO1 and HD, disrupted mitosis and prevented subsequent cell division. CHO1 is thought to exert this effect by blocking the activity of a 110-kD KLP. The relevant target of HD, which was raised against the KHC motor domain, is unknown; HD may disrupt mitosis by interfering with an essential spindle KLP but not with KHC itself, as preabsorption of HD with KHC did not alter its ability to block mitosis. These data indicate that some KLPs have essential mitotic functions in early sea urchin embryos but KHC itself does not.

scholarly journals A novel mammalian protein, p55CDC, present in dividing cells is associated with protein kinase activity and has homology to the Saccharomyces cerevisiae cell division cycle proteins Cdc20 and Cdc4.

1994 ◽  
Vol 14 (5) ◽  
pp. 3350-3363 ◽  
Author(s):  
J Weinstein ◽  
F W Jacobsen ◽  
J Hsu-Chen ◽  
T Wu ◽  
L G Baum
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Division ◽  
Protein Kinase ◽  
Mammalian Cells ◽  
Kinase Activity ◽  
Yeast Cells ◽  
Protein Kinase Activity ◽  
Growing Cell ◽  
Dividing Cells

A novel protein, p55CDC, has been identified in cycling mammalian cells. This transcript is readily detectable in all exponentially growing cell lines but disappears when cells are chemically induced to fall out of the cell cycle and differentiate. The p55CDC protein appears to be essential for cell division, since transfection of antisense p55CDC cDNA into CHO cells resulted in isolation of only those cells which exhibited a compensatory increase in p55CDC transcripts in the sense orientation. Immunoprecipitation of p55CDC yielded protein complexes with kinase activity which fluctuated during the cell cycle. Since p55CDC does not have the conserved protein kinase domains, this activity must be due to one or more of the associated proteins in the immune complex. The highest levels of protein kinase activity were seen with alpha-casein and myelin basic protein as substrates and demonstrated a pattern of activity distinct from that described for the known cyclin-dependent cell division kinases. The p55CDC protein was also phosphorylated in dividing cells. The amino acid sequence of p55CDC contains seven repeats homologous to the beta subunit of G proteins, and the highest degree of homology in these repeats was found with the Saccharomyces cerevisiae Cdc20 and Cdc4 proteins, which have been proposed to be involved in the formation of a functional bipolar mitotic spindle in yeast cells. The G beta repeat has been postulated to mediate protein-protein interactions and, in p55CDC, may modulate its association with a unique cell cycle protein kinase. These findings suggest that p55CDC is a component of the mammalian cell cycle mechanism.

Regulation of Microtubule Assembly-Disassembly in Mitotic Mammalian Cells: Role of Calcium, Calmodulin and MTOCs

1981 ◽  
Vol 39 ◽  
pp. 572-575
Author(s):  
B. R. Brinkley ◽  
S. L. Brenner ◽  
D. A. Pepper ◽  
R. L. Pardue
Keyword(s):  
Cell Cycle ◽  
Calcium Binding ◽  
Mammalian Cells ◽  
Microtubule Assembly ◽  
Free Calcium ◽  
Mitotic Apparatus ◽  
Cytoplasmic Microtubule ◽  
Dividing Cells ◽  
Late Telophase

Two microtubule arrays exist in cultured mammalian cells during their progression through the cell cycle; the cytoplasmic microtubule complexes (CMTC) of interphase cells (Figure 1) and the mitotic apparatus (MA) of dividing cells (Figure 2). As chromosomes are segregated to opposite poles of the spindle during telophase, the microtubules of the MA are disassembled. During late telophase -G1 phase the tubulin subunits from the spindle are recycled into the microtubules of the CMTC which forms an elaborate network throught the cytoplasm. When cells progress into late G2 -Prophase, the CMTC is disassembled and the tubulin is converted into microtubules of the MA. our research has been aimed at defining the mechanism whereby cells regulate the alternating patterns of microtubule assembly-disassembly during the cell cycle.In one series of experiments, we have investigated the role of calcium in microtubule assembly. Several laboratories have shown that cytoplasmic and spindle microtubules are unstable in the presence of elevated free calcium levels. Using monospecific antibodies and indirect immunofluorescence, we have demonstrated the presence of the ubiquitous calcium-binding protein calmodulin in the mitotic spindle of mammalian cells in vitro (Figure 3).

scholarly journals NuMA is a major acceptor of poly(ADP-ribosyl)ation by tankyrase 1 in mitosis

2005 ◽  
Vol 391 (2) ◽  
pp. 177-184 ◽  
Author(s):  
William Chang ◽  
Jasmin N. Dynek ◽  
Susan Smith
Keyword(s):  
Small Interfering Rna ◽  
Complete Loss ◽  
Mitotic Progression ◽  
Mitotic Apparatus ◽  
Spindle Poles ◽  
Early Anaphase ◽  
Parp Activity ◽  
Sirna Knockdown ◽  
Tankyrase 1 ◽  
Interfering Rna

Tankyrase 1 is a PARP [poly(ADP-ribose) polymerase] that localizes to multiple subcellular sites, including telomeres and mitotic centrosomes. Previous studies demonstrated that cells deficient in tankyrase 1 suffered a block in resolution of sister telomeres and arrested in early anaphase [Dynek and Smith (2004) Science 304, 97–100]. This phenotype was dependent on the catalytic PARP activity of tankyrase 1. To identify critical acceptors of PARsylation [poly(ADP-ribosyl)ation] by tankyrase 1 in mitosis, tankyrase 1 immunoprecipitates were analysed for associated PARsylated proteins. We identified NuMA (nuclear mitotic apparatus protein) as a major acceptor of poly(ADP-ribose) from tankyrase 1 in mitosis. We showed by immunofluorescence and immunoprecipitation that association between tankyrase 1 and NuMA increases dramatically at the onset of mitosis, concomitant with PARsylation of NuMA. Knockdown of tankyrase 1 by siRNA (small interfering RNA) eliminates PARsylation of NuMA in mitosis, confirming tankyrase 1 as the PARP responsible for this modification. However, even in the absence of tankyrase 1 and PARsylation, NuMA localizes to spindle poles. By contrast, siRNA knockdown of NuMA results in complete loss of tankyrase 1 from spindle poles. We discuss our result in terms of a model where PARsylation of NuMA by tankyrase 1 in mitosis could play a role in sister telomere separation and/or mitotic progression.

scholarly journals Kinase activation of ClC-3 accelerates cytoplasmic condensation during mitotic cell rounding

2012 ◽  
Vol 302 (3) ◽  
pp. C527-C538 ◽  
Author(s):  
Vishnu Anand Cuddapah ◽  
Christa W. Habela ◽  
Stacey Watkins ◽  
Lindsay S. Moore ◽  
Tia-Tabitha C. Barclay ◽  
...  
Keyword(s):  
Cell Division ◽  
Osmotic Pressure ◽  
Mammalian Cells ◽  
Shape Change ◽  
Time Lapse ◽  
Mitotic Cell ◽  
Cell Rounding ◽  
Gated Channel ◽  
Dividing Cells ◽  
Preceding Cell

“Mitotic cell rounding” describes the rounding of mammalian cells before dividing into two daughter cells. This shape change requires coordinated cytoskeletal contraction and changes in osmotic pressure. While considerable research has been devoted to understanding mechanisms underlying cytoskeletal contraction, little is known about how osmotic gradients are involved in cell division. Here we describe cytoplasmic condensation preceding cell division, termed “premitotic condensation” (PMC), which involves cells extruding osmotically active Cl− via ClC-3, a voltage-gated channel/transporter. This leads to a decrease in cytoplasmic volume during mitotic cell rounding and cell division. Using a combination of time-lapse microscopy and biophysical measurements, we demonstrate that PMC involves the activation of ClC-3 by Ca2+/calmodulin-dependent protein kinase II (CaMKII) in human glioma cells. Knockdown of endogenous ClC-3 protein expression eliminated CaMKII-dependent Cl− currents in dividing cells and impeded PMC. Thus, kinase-dependent changes in Cl− conductance contribute to an outward osmotic pressure in dividing cells, which facilitates cytoplasmic condensation preceding cell division.

A novel mammalian protein, p55CDC, present in dividing cells is associated with protein kinase activity and has homology to the Saccharomyces cerevisiae cell division cycle proteins Cdc20 and Cdc4

1994 ◽  
Vol 14 (5) ◽  
pp. 3350-3363
Author(s):  
J Weinstein ◽  
F W Jacobsen ◽  
J Hsu-Chen ◽  
T Wu ◽  
L G Baum
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Division ◽  
Protein Kinase ◽  
Mammalian Cells ◽  
Kinase Activity ◽  
Yeast Cells ◽  
Protein Kinase Activity ◽  
Growing Cell ◽  
Dividing Cells

A novel protein, p55CDC, has been identified in cycling mammalian cells. This transcript is readily detectable in all exponentially growing cell lines but disappears when cells are chemically induced to fall out of the cell cycle and differentiate. The p55CDC protein appears to be essential for cell division, since transfection of antisense p55CDC cDNA into CHO cells resulted in isolation of only those cells which exhibited a compensatory increase in p55CDC transcripts in the sense orientation. Immunoprecipitation of p55CDC yielded protein complexes with kinase activity which fluctuated during the cell cycle. Since p55CDC does not have the conserved protein kinase domains, this activity must be due to one or more of the associated proteins in the immune complex. The highest levels of protein kinase activity were seen with alpha-casein and myelin basic protein as substrates and demonstrated a pattern of activity distinct from that described for the known cyclin-dependent cell division kinases. The p55CDC protein was also phosphorylated in dividing cells. The amino acid sequence of p55CDC contains seven repeats homologous to the beta subunit of G proteins, and the highest degree of homology in these repeats was found with the Saccharomyces cerevisiae Cdc20 and Cdc4 proteins, which have been proposed to be involved in the formation of a functional bipolar mitotic spindle in yeast cells. The G beta repeat has been postulated to mediate protein-protein interactions and, in p55CDC, may modulate its association with a unique cell cycle protein kinase. These findings suggest that p55CDC is a component of the mammalian cell cycle mechanism.

Centromere assembly and non-random sister chromatid segregation in stem cells

2020 ◽  
Vol 64 (2) ◽  
pp. 223-232 ◽  
Author(s):  
Ben L. Carty ◽  
Elaine M. Dunleavy
Keyword(s):  
Stem Cells ◽  
Cell Division ◽  
Cell Fate ◽  
Sister Chromatid ◽  
Asymmetric Cell Division ◽  
Protein A ◽  
Germline Stem Cells ◽  
Sister Chromatids ◽  
Centromere Protein A ◽  
Oocyte Meiosis

Abstract Asymmetric cell division (ACD) produces daughter cells with separate distinct cell fates and is critical for the development and regulation of multicellular organisms. Epigenetic mechanisms are key players in cell fate determination. Centromeres, epigenetically specified loci defined by the presence of the histone H3-variant, centromere protein A (CENP-A), are essential for chromosome segregation at cell division. ACDs in stem cells and in oocyte meiosis have been proposed to be reliant on centromere integrity for the regulation of the non-random segregation of chromosomes. It has recently been shown that CENP-A is asymmetrically distributed between the centromeres of sister chromatids in male and female Drosophila germline stem cells (GSCs), with more CENP-A on sister chromatids to be segregated to the GSC. This imbalance in centromere strength correlates with the temporal and asymmetric assembly of the mitotic spindle and potentially orientates the cell to allow for biased sister chromatid retention in stem cells. In this essay, we discuss the recent evidence for asymmetric sister centromeres in stem cells. Thereafter, we discuss mechanistic avenues to establish this sister centromere asymmetry and how it ultimately might influence cell fate.

Sign in / Sign up

Export Citation Format

Share Document