scholarly journals Cell Cycle-regulated, Microtubule-independent Organelle Division in Cyanidioschyzon merolae

2005 ◽  
Vol 16 (5) ◽  
pp. 2493-2502 ◽  
Author(s):  
Keiji Nishida ◽  
Fumi Yagisawa ◽  
Haruko Kuroiwa ◽  
Toshiyuki Nagata ◽  
Tsuneyoshi Kuroiwa
Keyword(s):  
Cell Cycle ◽  
Spindle Pole ◽  
Chloroplast Division ◽  
Mitochondrial Morphology ◽  
Cyanidioschyzon Merolae ◽  
Microtubule Organization ◽  
Mitochondrial Division ◽  
Protein Levels ◽  
Division Site ◽  
Organelle Division

Mitochondrial and chloroplast division controls the number and morphology of organelles, but how cells regulate organelle division remains to be clarified. Here, we show that each step of mitochondrial and chloroplast division is closely associated with the cell cycle in Cyanidioschyzon merolae. Electron microscopy revealed direct associations between the spindle pole bodies and mitochondria, suggesting that mitochondrial distribution is physically coupled with mitosis. Interconnected organelles were fractionated under microtubule-stabilizing condition. Immunoblotting analysis revealed that the protein levels required for organelle division increased before microtubule changes upon cell division, indicating that regulation of protein expression for organelle division is distinct from that of cytokinesis. At the mitochondrial division site, dynamin stuck to one of the divided mitochondria and was spatially associated with the tip of a microtubule stretching from the other one. Inhibition of microtubule organization, proteasome activity or DNA synthesis, respectively, induced arrested cells with divided but shrunk mitochondria, with divided and segregated mitochondria, or with incomplete mitochondrial division restrained at the final severance, and repetitive chloroplast division. The results indicated that mitochondrial morphology and segregation but not division depend on microtubules and implied that the division processes of the two organelles are regulated at distinct checkpoints.

scholarly journals Chloroplast division checkpoint in eukaryotic algae

2016 ◽  
Vol 113 (47) ◽  
pp. E7629-E7638 ◽  
Author(s):  
Nobuko Sumiya ◽  
Takayuki Fujiwara ◽  
Atsuko Era ◽  
Shin-ya Miyagishima
Keyword(s):  
Cell Cycle ◽  
Host Cell ◽  
Cell Cycle Progression ◽  
Chloroplast Division ◽  
Dominant Negative ◽  
Cyanidioschyzon Merolae ◽  
Temperature Sensitive ◽  
Specific Expression ◽  
Cycle Progression ◽  
Division Site

Chloroplasts evolved from a cyanobacterial endosymbiont. It is believed that the synchronization of endosymbiotic and host cell division, as is commonly seen in existing algae, was a critical step in establishing the permanent organelle. Algal cells typically contain one or only a small number of chloroplasts that divide once per host cell cycle. This division is based partly on the S-phase–specific expression of nucleus-encoded proteins that constitute the chloroplast-division machinery. In this study, using the red alga Cyanidioschyzon merolae, we show that cell-cycle progression is arrested at the prophase when chloroplast division is blocked before the formation of the chloroplast-division machinery by the overexpression of Filamenting temperature-sensitive (Fts) Z2-1 (Fts72-1), but the cell cycle progresses when chloroplast division is blocked during division-site constriction by the overexpression of either FtsZ2-1 or a dominant-negative form of dynamin-related protein 5B (DRP5B). In the cells arrested in the prophase, the increase in the cyclin B level and the migration of cyclin-dependent kinase B (CDKB) were blocked. These results suggest that chloroplast division restricts host cell-cycle progression so that the cell cycle progresses to the metaphase only when chloroplast division has commenced. Thus, chloroplast division and host cell-cycle progression are synchronized by an interactive restriction that takes place between the nucleus and the chloroplast. In addition, we observed a similar pattern of cell-cycle arrest upon the blockage of chloroplast division in the glaucophyte alga Cyanophora paradoxa, raising the possibility that the chloroplast division checkpoint contributed to the establishment of the permanent organelle.

scholarly journals Distinct Dgrip84 Isoforms Correlate with Distinct γ-Tubulins in Drosophila

2008 ◽  
Vol 19 (1) ◽  
pp. 368-377 ◽  
Author(s):  
Christiane Wiese
Keyword(s):  
Cell Cycle ◽  
Spindle Pole ◽  
Multiprotein Complex ◽  
Microtubule Organization ◽  
Developmentally Regulated ◽  
Tubulin Genes ◽  
Tubulin Isotype ◽  
Ring Complex ◽  
Cycle Dependent ◽  
Drosophila Embryos

γ-Tubulin is an indispensable component of the animal centrosome and is required for proper microtubule organization. Within the cell, γ-tubulin exists in a multiprotein complex containing between two (some yeasts) and six or more (metazoa) additional highly conserved proteins named gamma ring proteins (Grips) or gamma complex proteins (GCPs). γ-Tubulin containing complexes isolated from Xenopus eggs or Drosophila embryos appear ring-shaped and have therefore been named the γ-tubulin ring complex (γTuRC). Curiously, many organisms (including humans) have two distinct γ-tubulin genes. In Drosophila, where the two γ-tubulin isotypes have been studied most extensively, the γ-tubulin genes are developmentally regulated: the “maternal” γ-tubulin isotype (named γTub37CD according to its location on the genetic map) is expressed in the ovary and is deposited in the egg, where it is thought to orchestrate the meiotic and early embryonic cleavages. The second γ-tubulin isotype (γTub23C) is ubiquitously expressed and persists in most of the cells of the adult fly. In those rare cases where both γ-tubulins coexist in the same cell, they show distinct subcellular distributions and cell-cycle-dependent changes: γTub37CD mainly localizes to the centrosome, where its levels vary only slightly with the cell cycle. In contrast, the level of γTub23C at the centrosome increases at the beginning of mitosis, and γTub23C also associates with spindle pole microtubules. Here, we show that γTub23C forms discrete complexes that closely resemble the complexes formed by γTub37CD. Surprisingly, however, γTub23C associates with a distinct, longer splice variant of Dgrip84. This may reflect a role for Dgrip84 in regulating the activity and/or the location of the γ-tubulin complexes formed with γTub37CD and γTub23C.

scholarly journals Glycosyltransferase MDR1 assembles a dividing ring for mitochondrial proliferation comprising polyglucan nanofilaments

2017 ◽  
Vol 114 (50) ◽  
pp. 13284-13289 ◽  
Author(s):  
Yamato Yoshida ◽  
Haruko Kuroiwa ◽  
Takashi Shimada ◽  
Masaki Yoshida ◽  
Mio Ohnuma ◽  
...  
Keyword(s):  
Skeletal Structure ◽  
Cyanidioschyzon Merolae ◽  
Free Living ◽  
Mitochondrial Division ◽  
Division Site ◽  
Ring Assembly ◽  
Assembly Mechanism ◽  
Single Ring ◽  
Binary Division ◽  
Nanoscale Imaging

Mitochondria, which evolved from a free-living bacterial ancestor, contain their own genomes and genetic systems and are produced from preexisting mitochondria by binary division. The mitochondrion-dividing (MD) ring is the main skeletal structure of the mitochondrial division machinery. However, the assembly mechanism and molecular identity of the MD ring are unknown. Multi-omics analysis of isolated mitochondrial division machinery from the unicellular alga Cyanidioschyzon merolae revealed an uncharacterized glycosyltransferase, MITOCHONDRION-DIVIDING RING1 (MDR1), which is specifically expressed during mitochondrial division and forms a single ring at the mitochondrial division site. Nanoscale imaging using immunoelectron microscopy and componential analysis demonstrated that MDR1 is involved in MD ring formation and that the MD ring filaments are composed of glycosylated MDR1 and polymeric glucose nanofilaments. Down-regulation of MDR1 strongly interrupted mitochondrial division and obstructed MD ring assembly. Taken together, our results suggest that MDR1 mediates the synthesis of polyglucan nanofilaments that assemble to form the MD ring. Given that a homolog of MDR1 performs similar functions in chloroplast division, the establishment of MDR1 family proteins appears to have been a singular, crucial event for the emergence of endosymbiotic organelles.

scholarly journals Fission Yeast mto2p Regulates Microtubule Nucleation by the Centrosomin-related Protein mto1p

2005 ◽  
Vol 16 (6) ◽  
pp. 3040-3051 ◽  
Author(s):  
Itaru Samejima ◽  
Paula C. C. Lourenço ◽  
Hilary A. Snaith ◽  
Kenneth E. Sawin
Keyword(s):  
Fission Yeast ◽  
Insertional Mutagenesis ◽  
Spindle Pole ◽  
Related Protein ◽  
Microtubule Nucleation ◽  
Microtubule Organization ◽  
Cytoplasmic Microtubule ◽  
Division Site ◽  
Cell Extracts

From an insertional mutagenesis screen, we isolated a novel gene, mto2+, involved in microtubule organization in fission yeast. mto2Δ strains are viable but exhibit defects in interphase microtubule nucleation and in formation of the postanaphase microtubule array at the end of mitosis. The mto2Δ defects represent a subset of the defects displayed by cells deleted for mto1+ (also known as mod20+ and mbo1+), a centrosomin-related protein required to recruit the γ-tubulin complex to cytoplasmic microtubule-organizing centers (MTOCs). We show that mto2p colocalizes with mto1p at MTOCs throughout the cell cycle and that mto1p and mto2p coimmunoprecipitate from cytoplasmic extracts. In vitro studies suggest that mto2p binds directly to mto1p. In mto2Δ mutants, although some aspects of mto1p localization are perturbed, mto1p can still localize to spindle pole bodies and the cell division site and to “satellite” particles on interphase microtubules. In mto1Δ mutants, localization of mto2p to all of these MTOCs is strongly reduced or absent. We also find that in mto2Δ mutants, cytoplasmic forms of the γ-tubulin complex are mislocalized, and the γ-tubulin complex no longer coimmunoprecipitates with mto1p from cell extracts. These experiments establish mto2p as a major regulator of mto1p-mediated microtubule nucleation by the γ-tubulin complex.

scholarly journals Phosphorylation in the intrinsically disordered region of F-BAR protein Imp2 regulates its contractile ring recruitment

2021 ◽  
Author(s):  
Alaina H. Willet ◽  
Maya G. Igarashi ◽  
Jun-Song Chen ◽  
Rahul Bhattacharjee ◽  
Liping Ren ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Contractile Ring ◽  
Casein Kinase 1 ◽  
Protein Levels ◽  
Intrinsically Disordered ◽  
Division Site ◽  
Protein Partners ◽  
Intrinsically Disordered Region ◽  
And Function

The F-BAR protein Imp2 is an important contributor to cytokinesis in the fission yeast, Schizosaccharomyces pombe. Because cell cycle regulated phosphorylation of the central intrinsically disordered region (IDR) of the Imp2 paralog, Cdc15, controls Cdc15 oligomerization state, localization, and ability to bind protein partners, we investigated whether Imp2 is similarly phosphoregulated. We found that Imp2 is endogenously phosphorylated on 28 sites within its IDR with the bulk of phosphorylation being constitutive. In vitro, casein kinase 1 (CK1) Hhp1 and Hhp2 can phosphorylate 17 sites and Cdk1 the remaining 11 sites. Mutations that prevent Cdk1 phosphorylation result in precocious Imp2 recruitment to the cell division site, and mutations designed to mimic these phosphorylation events delay Imp2 CR accumulation. Mutations that eliminated CK1 phosphorylation sites allowed CR sliding, and phosphomimetic substitutions at these sites reduced Imp2 protein levels and slowed CR constriction. Thus, like Cdc15, the Imp2 IDR is phosphorylated at many sites by multiple kinases. In contrast to Cdc15, for which phosphorylation plays a major cell cycle regulatory role, Imp2 phosphorylation is primarily constitutive with milder effects on localization and function.

scholarly journals Microtubule Organization Requires Cell Cycle-dependent Nucleation at Dispersed Cytoplasmic Sites: Polar and Perinuclear Microtubule Organizing Centers in the Plant Pathogen Ustilago maydis

2003 ◽  
Vol 14 (2) ◽  
pp. 642-657 ◽  
Author(s):  
Anne Straube ◽  
Marianne Brill ◽  
Berl R. Oakley ◽  
Tetsuya Horio ◽  
Gero Steinberg
Keyword(s):  
Cell Cycle ◽  
Fluorescent Protein ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Microtubule Organization ◽  
Microtubule Organizing Centers ◽  
Nucleation Sites ◽  
Cycle Dependent ◽  
Green Fluorescent

Growth of most eukaryotic cells requires directed transport along microtubules (MTs) that are nucleated at nuclear-associated microtubule organizing centers (MTOCs), such as the centrosome and the fungal spindle pole body (SPB). Herein, we show that the pathogenic fungusUstilago maydis uses different MT nucleation sites to rearrange MTs during the cell cycle. In vivo observation of green fluorescent protein-MTs and MT plus-ends, tagged by a fluorescent EB1 homologue, provided evidence for antipolar MT orientation and dispersed cytoplasmic MT nucleating centers in unbudded cells. On budding γ-tubulin containing MTOCs formed at the bud neck, and MTs reorganized with >85% of all minus-ends being focused toward the growth region. Experimentally induced lateral budding resulted in MTs that curved out of the bud, again supporting the notion that polar growth requires polar MT nucleation. Depletion or overexpression of Tub2, the γ-tubulin from U. maydis, affected MT number in interphase cells. The SPB was inactive in G2 phase but continuously recruited γ-tubulin until it started to nucleate mitotic MTs. Taken together, our data suggest that MT reorganization in U. maydis depends on cell cycle-specific nucleation at dispersed cytoplasmic sites, at a polar MTOC and the SPB.

scholarly journals An anillin homologue, Mid2p, acts during fission yeast cytokinesis to organize the septin ring and promote cell separation

2003 ◽  
Vol 160 (7) ◽  
pp. 1093-1103 ◽  
Author(s):  
Joseph J. Tasto ◽  
Jennifer L. Morrell ◽  
Kathleen L. Gould
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Fission Yeast ◽  
Cell Separation ◽  
Cell Cycle Progression ◽  
Dependent Manner ◽  
Septin Ring ◽  
Protein Levels ◽  
Division Site ◽  
Cell Biol

Anillin is a conserved protein required for cell division (Field, C.M., and B.M. Alberts. 1995. J. Cell Biol. 131:165–178; Oegema, K., M.S. Savoian, T.J. Mitchison, and C.M. Field. 2000. J. Cell Biol. 150:539–552). One fission yeast homologue of anillin, Mid1p, is necessary for the proper placement of the division site within the cell (Chang, F., A. Woollard, and P. Nurse. 1996. J. Cell Sci. 109(Pt 1):131–142; Sohrmann, M., C. Fankhauser, C. Brodbeck, and V. Simanis. 1996. Genes Dev. 10:2707–2719). Here, we identify and characterize a second fission yeast anillin homologue, Mid2p, which is not orthologous with Mid1p. Mid2p localizes as a single ring in the middle of the cell after anaphase in a septin- and actin-dependent manner and splits into two rings during septation. Mid2p colocalizes with septins, and mid2Δ cells display disorganized, diffuse septin rings and a cell separation defect similar to septin deletion strains. mid2 gene expression and protein levels fluctuate during the cell cycle in a sep1- and Skp1/Cdc53/F-box (SCF)–dependent manner, respectively, implying that Mid2p activity must be carefully regulated. Overproduction of Mid2p depolarizes cell growth and affects the organization of both the septin and actin cytoskeletons. In the presence of a nondegradable Mid2p fragment, the septin ring is stabilized and cell cycle progression is delayed. These results suggest that Mid2p influences septin ring organization at the site of cell division and its turnover might normally be required to permit septin ring disassembly.

Variation in cytoplasmic microtubule organization and spindle length between the two forms of the dimorphic fungus Candida albicans

1988 ◽  
Vol 91 (2) ◽  
pp. 211-220
Author(s):  
R. Barton ◽  
K. Gull
Keyword(s):  
Cell Cycle ◽  
Candida Albicans ◽  
Mitotic Spindle ◽  
Spindle Pole ◽  
Yeast Cells ◽  
Dimorphic Fungus ◽  
Microtubule Organization ◽  
Cytoplasmic Microtubule ◽  
Yeast Phase ◽  
Cytoplasmic Microtubules

Candida albicans is a dimorphic fungus capable of growing as a budding yeast and as a filamentous hypha. We have used the technique of immunofluorescence to study the changes in the microtubule cytoskeleton during the cell cycle in both growth forms. This approach has revealed the presence of an extensive system of microtubules, including cytoplasmic microtubules and a rod-like intranuclear spindle. We have provided a complete description of the arrangement of cytoplasmic and spindle microtubules at each phase of the yeast cell cycle. A novel and characteristic feature of the yeast phase of Candida is the presence of an array of short microtubules at the neck of the doublet cell. This neck-associated array (NAA), is apparently organized independently of the main microtubule-organizing centre, the spindle pole bodies, at late anaphase. Analysis of microtubule patterns in the hyphal state reveals that the general arrangements of microtubules are similar to those seen in the yeast phase. These patterns suggest a role for the cytoplasmic microtubules in the nuclear migration that occurs during hyphal growth. A major finding is that the mitotic spindle in hyphae is considerably longer (12–20 microns) than the spindle in yeast cells (7–8 microns). This may reflect the role of the hyphal mitotic spindle not only in nuclear division but also in the positioning of the daughter nuclei at the centres of hyphal compartments.

scholarly journals Two Dictyostelium Orthologs of the Prokaryotic Cell Division Protein FtsZ Localize to Mitochondria and Are Required for the Maintenance of Normal Mitochondrial Morphology

2003 ◽  
Vol 2 (6) ◽  
pp. 1315-1326 ◽  
Author(s):  
Paul R. Gilson ◽  
Xuan-Chuan Yu ◽  
Dale Hereld ◽  
Christian Barth ◽  
Amelia Savage ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Higher Plants ◽  
Principal Component ◽  
Mitochondrial Morphology ◽  
Wild Type ◽  
Mitochondrial Division ◽  
Organelle Division ◽  
Cell Division Protein ◽  
The One ◽  
Green Fluorescent

ABSTRACT In bacteria, the protein FtsZ is the principal component of a ring that constricts the cell at division. Though all mitochondria probably arose through a single, ancient bacterial endosymbiosis, the mitochondria of only certain protists appear to have retained FtsZ, and the protein is absent from the mitochondria of fungi, animals, and higher plants. We have investigated the role that FtsZ plays in mitochondrial division in the genetically tractable protist Dictyostelium discoideum, which has two nuclearly encoded FtsZs, FszA and FszB, that are targeted to the inside of mitochondria. In most wild-type amoebae, the mitochondria are spherical or rod-shaped, but in fsz-null mutants they become elongated into tubules, indicating that a decrease in mitochondrial division has occurred. In support of this role in organelle division, antibodies to FszA and FszA-green fluorescent protein (GFP) show belts and puncta at multiple places along the mitochondria, which may define future or recent sites of division. FszB-GFP, in contrast, locates to an electron-dense, submitochondrial body usually located at one end of the organelle, but how it functions during division is unclear. This is the first demonstration of two differentially localized FtsZs within the one organelle, and it points to a divergence in the roles of these two proteins.

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