Nuclear DNA,nuclear area and nuclear dry mass in thirteen species of Crotalaria (Angiospermae,Leguminosae)

Estimations on nuclear DNA content, nuclear area and nuclear dry mass were made on 13 species of Crotalaria, with the help of Vicker's M 85 microdensitometer and Vicker's interferometer. Highly significant differences for all three characters were found between species. DNA density (DNA/area) increased with increase in DNA content. The cultivated species, C. juncea had a lower DNA density relative to its DNA content.—Nuclear DNA made only a small fraction of nuclear dry mass (115). Although total dry mass and non-nucleolar nuclear dry mass showed significant regression on DNA content, the nucleolar dry mass gave a much wider scatter, indicating that nucleolar mass is not equally dependent on nuclear DNA content.     

Les ultrastructures nucléaires de la Noctiluque (Dinoflagellé libre) au cours de la sporogenèse

The trophozoït of Noctiluca miliaris has a large nucleus (30 ) with several nucleoli of considerable size that contain DNA fibrillae lying in the interspaces. — Before and during the first sporogenetic divisions, the nucleoli disintegrate, releasing towards the cytoplasma numerous groups of ribonucleic granules passing through the nuclear ampullae. At the end of the sporulation, there are no nucleoli visible in the nuclei and no ampullae. — The nucleoplasm diminishes, as the DNA filaments are built up, to form the meshes of a network which limit the masses of chromatic material that take the shape of chromosomes characterized by regular fibrillar arches, at the 8–16 nuclei stage. In their centre, there is an axial structure which remains intact during the chromosomal segregation; its function during mitosis seems to be important: supplementary layers of arches appear at this level. — The progressive condensation of the chromosomes is correlated to the sporogenetic evolution of the nuclei, not to the different phases of the mitotic cycle. — The karyokinesis is brought about, during early stages, by mere splitting of the chromatic mass and of its envelope, and later one by separation into two lots of chromosomes. The segregation of these chromosomes is effected by partial intervention and growth of the envelope of the nucleus; there is no centromeric structure visible. At the end of divisions, the nucleus is almost entirely formed by its chromosomes. — The nucleolar structure, the karyokinesis, the structure of the nuclear envelope and the chromosomal cycle show the particularly high evolution of Noctiluca, within the Dinoflagellata.     

Subnuclear redistribution of DNA species in confluent and growing mammalian cells

About 20 to 25 percent of the nuclear DNA from cultured cells of the African green monkey, Cercopithecus aethiops, consists of a homogeneous, highly repetitive fraction designated C. aethiops component DNA. Use of in situ hybridization techniques reveals component at the centromeres of chromosomes from both diploid and heteroploid African green monkey kidney (AGMK) tissue culture cells. — Component DNA comprises 47 percent of the nucleolar DNA in actively growing primary AGMK cells, but only 31 percent of the nucleolar DNA in confluent cells which show density-dependent growth inhibition. Further, there is a pronounced shift of both main band and component DNA from euchromatin to heterochromatin when growing cells attain confluency. Thus, the relative subnuclear distributions of component and main band DNA's are different in growing and confluent cells. — In situ hybridization techniques indicate that component sequences aggregate in clumps in nuclei of growing cells and show a diffuse distribution in nuclei of confluent cells. This suggests that centromeric regions of the various chromosomes or groups of chromosomes aggregate and disaggregate reversibly as the culture changes from density-dependent growth inhibition to active cell division. — Hypotonic citrate treatment of primary AGMK cells causes nucleoli of confluent cells to disperse: this dispersion following citrate treatment was not seen in growing AGMK cells or in confluent or growing heteroploid cells. Similarly, this nucleolar dispersion was seen in confluent diploid mouse and human cells but not in growing diploid cells or in confluent or growing heteroploid cells.     

Mitosis in the coenocytic green algaBoergesenia forbesii (Harvey) Feldmann (Siphonocladales,Ulvophyceae)

Mitosis in vegetative cells of the siphonocladalean algaBoergesenia forbesii (Harvey) Feldmann was investigated mainly by electron microscopy. The mitotic spindle was centric and closed. The interphase nucleus contained a spherical nucleolus. The nucleolus was slightly dispersed at prophase, but nucleolar materials remained during nearly all stages of mitosis. Kinetochores were evident on chromosomes. The polar regions of nuclear envelope had no fenestrae during mitosis. Anaphase separation of the chromosomes was asynchronous. Elongation of interzonal spindle at telophase separated the two daughter nuclei widely. The ultrastructural features of mitosis inB. forbesii revealed by the present investigation are compared with those of other siphonous and siphonocladous algae in the Ulvophyceae.     

Nucleolar localization and identification of nuclear/nucleolar localization signals of the calmodulin-binding protein nucleomorphin during growth and mitosis in <Emphasis Type="Italic">Dictyostelium</Emphasis>

The calmodulin-binding protein nucleomorphin isoform NumA1 is a nuclear number regulator in Dictyostelium that localizes to intra-nuclear patches adjacent to the nuclear envelope and to a lesser extent the nucleoplasm. Earlier studies have shown similar patches to be nucleoli but only three nucleolar proteins have been identified in Dictyostelium. Here, actinomycin-D treatment caused the loss of NumA1 localization, while calcium and calmodulin antagonists had no effect. In keeping with a nucleolar function, NumA1 moved out of the presumptive nucleoli during mitosis redistributing to areas within the nucleus, the spindle fibers, and centrosomal region before re-accumulating in the presumptive nucleoli at telophase. Together, these data verify NumA1 as a true nucleolar protein. Prior to this study, the dynamics of specific nucleolar proteins had not been determined during mitosis in Dictyostelium. FITC-conjugated peptides equivalent to presumptive nuclear localization signals within NumA1 localized to nucleoli indicating that they also act as nucleolar localization signals. To our knowledge, these represent the first precisely defined nucleolar localization signals as well as the first nuclear/nucleolar localization signals identified in Dictyostelium. Together, these results reveal that NumA1 is a true nucleolar protein and the only nucleolar calmodulin-binding protein identified in Dictyostelium. The possible use of nuclear/nucleolar localization signal-mediated drug targeting to nucleoli is discussed.     

Yeast Srp1, a nuclear protein related toDrosophila and mouse pendulin, is required for normal migration, division, and integrity of nuclei during mitosis

This paper describes genes from yeast and mouse with significant sequence similarities to aDrosophila gene that encodes the blood cell tumor suppressor pendulin. The protein encoded by the yeast gene, Srp1p, and mouse pendulin share 42% and 51% amino acid identity withDrosophila pendulin, respectively. All three proteins consist of 10.5 degenerate tandem repeats of 42 amino acids each. Similar repeats occur in a superfamily of proteins that includes theDrosophila Armadillo protein. All three proteins contain a consensus sequence for a bipartite nuclear localization signal (NLS) in the N-terminal domain, which is not part of the repeat structure. Confocal microscopic analysis of yeast cells stained with antibodies against Srp1p reveals that this protein is intranuclear throughout the cell cycle. Targeted gene disruption shows thatSRP1 is an essential gene. Despite their sequence similarities,Drosophila and mouse pendulin are unable to rescue the lethality of anSRP1 disruption. We demonstrate that yeast cells depleted of Srp1p arrest in mitosis with a G2 content of DNA. Arrested cells display abnormal structures and orientations of the mitotic spindles, aberrant segregation of the chromatin and the nuclei, and threads of chromatin emanating from the bulk of nuclear DNA. This phenotype suggests that Srplp is required for the normal function of microtubules and the spindle pole bodies, as well as for nuclear integrity. We suggest that Srp1p interacts with multiple components of the cell nucleus that are required for mitosis and discuss its functional similarities to, and differences fromDrosophila pendulin.     

A nucleolar auto-antigen is part of a major chromosomal surface component

Several nucleolar antigens are defined by human autoantibodies. These antigens can therefore be used to follow the fate of nucleolar components through mitosis when this major nuclear structure disintegrates and becomes reassembled in G₁-phase. We found that fibrillarin leaves the nucleolus before complete breakdown of this structure and attaches to chromosomes before nuclear envelope breakdown. In mouse, fibrillarin attaches over the chromosomal surface except for the excluded centromeric region. The antigen is transported to the new nucleus via the chromosomes and is last seen on chromosomal surfaces facing the cytoplasm during nuclear envelope reformation. Lamin B reappears on the same chromosomal surfaces before the nucleolar antigen is removed and aggregates for new nucleolar reformation in G₁-phase cells. From our observations, we postulate that the antigen acts in concert with other proteins as a nuclear envelope equivalent by forming a protective sheath around the chromosome, that it excludes larger molecules, and helps to separate the chromosomes, in addition to segregation of the ribonucleoprotein (RNP) back to the nucleus for nucleolar reconstruction. We also suggest that the selective retention of these antigens from certain areas on individual chromosomes together with specific lamin B attachment over these chromosomal surfaces allows for a nonrandom positioning of chromosomes in the nucleus.     

Protein Phosphorylation During the Transitions from G1 Arrest Point to S and G2 Arrest Point to M in Vicia faba Root Meristem

Using monoclonal FITC conjugated antibodies that specifically recognize phosphorylated form of threonine (-T^P-FITC) it was shown that in excised, sugar-starved root meristems of Vicia faba subsp. minor the expression of two principal control points (PCPs) is correlated with a marked decrease in nuclear and nucleolar phosphorylation of proteins. When stationary phase meristems are supplied with 2 % sucrose, the G1-arrested cells start out DNA replication (assessed by pulse ³H-thymidine labeling), while the cells blocked in G2 phase initiate mitotic divisions. Using this model, we have found that depending on the period during which the sucrose-mediated PCP1S and PCP2M transitions are affected by 6-dimethylaminopurine, an inhibitor of protein kinases, the release from G1 and G2 phase arrest-points of the cell cycle become prolonged, showing different time-course modifications. Furthermore, our results seem to implicate that both PCP1 and PCP2 comprise subpopulations of cells differing with respect to the rates at which they commence DNA replication and mitosis.     

Synaptonemal complexes of normal and mutant yeast chromosomes (Saccharomyces cerevisiae)

Synaptonemal complex (SC) analysis of six laboratory yeast strains showed the SC karyotypes to be repeatable within strains. Chromosomal differences were found between strains. In five of the strains, two SCs insert into the nucleolus. This represents a single bivalent with a nucleolar organizer in a medial position as is suggested by genetic data or two bivalents each with a terminal nucleolar organizer. In the first interpretation, n=16; in the second, n=17. Strain Tris has a single nucleolar SC and n=17. In strains DCx374, DCx416 and x 8366a the genetically determined rearrangements of linkage group III could not be identified. Presumably the short SC (0.33 m) associated with linkage group III cannot accommodate an inversion loop or a translocation configuration. The strains however were found to harbour a reciprocal translocation involving the nucleolar chromosome. Trisomy for one of the longer chromosomes was observed in Tris and spo10. It is concluded that rearrangements of the medium and long but not short yeast chromosomes can be detected cytologically. — Measurements of nuclear volumes show SC length to vary with artifactually induced swelling of the nucleus. Linear regression of SC length over nuclear radius indicates that actual SC length is only about one-half the observed length. As a result the DNA packing per SC unit length is higher then previously estimated.     

Cytophotometric study of nuclear differentiation during pollen development in Tradescantia Paludosa

Summary During pollen development the dry weight, total protein, histone, DNA, arginine, and lysine content were analysed by cytophotometric methods in partially isolated nuclei. The amount of analysed substances increased from the end of the meiosis to the mitosis of the microspores to the double of the initial values. After mitosis the ratio histone/DNA remained almost unchanged in both vegetative and generative nuclei. On the other hand a large difference in the ratio non-histone protein/DNA could be observed, the vegetative nucleus containing more non-histone protein than the generative nucleus. The rate of RNA synthesis being higher in the vegetative nuclei, these non-histone proteins may have some function in nuclear activation. The DNA of the generative nucleus is duplicated before anthesis, whilst in the vegetative nucleus the DNA content remains constant.     

Mitosis in the fungusBasidiobolus ranarum as revealed by electron microscopy

Summary Mitosis of nuclei in vegetative hyphae of the fungusBasidiobolus ranarum has been studied by electron microscopy. Cells fixed with glutaraldehyde and OsO⁴ were embedded in Vestopal. Sections were obtained of single cells whose mitotic status was known. Attention was paid to the behaviour of the microtubules, the nuclear envelope and the nucleolus. Nuclear division begins with the dilution and rearrangement of nucleolar material and the gradual breakdown of the nuclear envelope. At this stage the nucleus is surrounded by a sheet of closely packed microtubules. Some of these penetrate into the nucleus through gaps in the envelope. Dissolution of the envelope is followed or accompanied by the development of an extensive labyrinth of membranous cisternae which persists at the periphery of the division site through mitosis and probably contributes material to the envelopes of the daughter nuclei. The drum-shaped spindle of metaphase is composed of large numbers of microtubules aligned parallel to each other. Many of them are associated with chromosomes. Metaphase is soon followed by the movement of dense masses of nucleolar material and chromosomes to the poles of the division figure to form the socalled end plates. Microtubules extend into the end plates but not beyond. Neither centrioles nor centriolar plaques have been seen.     

Rad53 homologue forkhead-associated kinase A (FhkA) and Ca<Superscript>2+</Superscript>-binding protein 4a (CBP4a) are nucleolar proteins that differentially redistribute during mitosis in <Emphasis Type="Italic">Dictyostelium</Emphasis>

Background

During mitosis most nucleolar proteins redistribute to other locales providing an opportunity to study the relationship between nucleolar protein localization and function. Dictyostelium is a model organism for the study of several fundamental biological processes and human diseases but only two nucleolar proteins have been studied during mitosis: NumA1 and Snf12. Both of them are linked to the cell cycle. To acquire a better understanding of nucleolar protein localization and dynamics in Dictyostelium we studied the nucleolar localization of two additional proteins during mitosis: Snf12-linked forkhead-associated kinase A (FhkA), which is involved in the cell cycle, and Ca²⁺-binding protein 4a (CBP4a), which is a binding partner of NumA1.

Methods

Polyclonal antibodies were produced in-house. Cells were fixed and probed with either anti-FhkA or anti-CBP4a in order to determine cellular localization during interphase and throughout the stages of mitosis. Colocalization with DAPI nuclear stain allowed us to determine the location of the nucleus and nucleolus while colocalization with anti-α-tubulin allowed us to determine the cell cycle stage.

Results

Here we verify two novel nucleolar proteins, Rad53 homologue FhkA which localized around the edge of the nucleolus and CBP4a which was detected throughout the entire nucleolus. Treatment with the Ca²⁺ chelator BAPTA (5?mM) showed that the nucleolar localization of CBP4a is Ca²⁺-dependent. In response to actinomycin D (0.05?mg/mL) CBP4a disappeared from the nucleolus while FhkA protruded from the nucleus, eventually pinching off as cytoplasmic circles. FhkA and CBP4a redistributed differently during mitosis. FhkA redistributed throughout the entire cell and at the nuclear envelope region from prometaphase through telophase. In contrast, during prometaphase CBP4a relocated to many large, discrete “CBP4a islands” throughout the nucleoplasm. Two larger “CBP4a islands” were also detected specifically at the metaphase plate region.

Conclusions

FhkA and CBP4a represent the sixth and seventh nucleolar proteins that have been verified to date in Dictyostelium and the third and fourth studied during mitosis. The protein-specific distributions of all of these nucleolar proteins during interphase and mitosis provide unique insight into nucleolar protein dynamics in this model organism setting the stage for future work.

     

Dynamic Changes in Nuclear Architecture during Mitosis: On the Role of Protein Phosphorylation in Spindle Assembly and Chromosome Segregation

During mitosis, the vertebrate cell nucleus undergoes profound changes in architecture. At the onset of mitosis, the nuclear envelope breaks down, the nuclear lamina is depolymerized, and interphase chromatin is condensed to chromosomes. Concomitantly, cytoplasmic microtubules are reorganized into a mitotic spindle apparatus, a highly dynamic structure required for the segregation of sister chromatids. Many of the above events are controlled by reversible phosphorylation. Hence, our laboratory is interested in characterizing the kinases involved in promoting progression through mitosis and in identifying their relevant substrates. Prominent among the kinases responsible for regulating entry into mitosis is the Cdc2 kinase, the first member of the cyclin dependent kinase (Cdk) family. Recently, we found that Cdc2 phosphorylates HsEg5, a human kinesin-related motor protein associated with centrosomes and the spindle apparatus. Our results indicate that phosphorylation regulates the association of HsEg5 with the mitotic spindle and that the function of this plus-end directed motor is essential for centrosome separation and bipolar spindle formation. Another kinase implicated in regulating progression through mitosis is Plk1 (polo-like kinase 1), the human homologue of theDrosophilagene product “polo.” By antibody microinjection we have found that Plk1 is required for the functional maturation of centrosomes and hence for entry into mitosis. Furthermore, we found that microinjected anti-Plk1 antibodies caused a more severe block to cell cycle progression in diploid fibroblasts than in immortalized tumor cells. This observation hints at the existence of a checkpoint linking Cdc2 activation to the presence of functional centrosomes.     

Estrogens and cell death in murine uterine luminal epithelium

Summary The luminal epithelium of adult ovariectomized mice responds to estradiol-17 with a synchronised wave of DNA synthesis and mitosis. Estriol, however, although producing a similar DNA-synthetic and mitotic response fails to cause an increase in cell number owing to a wave of cell death occurring at mitosis. In the present study it was shown that cells died by two different routes. The majority died by apoptosis but, unusually, a minority also died by necrosis. In the apoptotic cells the cytoplasm became dense, the endoplasmic reticulum and nuclear cisternae dilated; chromatin became marginated the nucleus shrank and became deeply infolded and contorted. Apoptosis, however, was uncharacteristic in that the nucleus failed to fragment, form caps or show disruption before the cells died by membrane rupture. Furthermore, the cells were frequently lost in sheets from the epithelium into the lumen. Part of the biochemical explanation for this onset of cell death comes from the accelerated loss from the tissue of estriol when compared to estradiol-17. This resulted in a decline in protein and rRNA biosynthesis and a failure to complete ribosomal maturation. Evidence in favour of this explanation came from experiments that showed a return to the estradiol-17 level of response and an inhibition of cell death when the occupancy of the estriol receptor was maintained.     

Identification and characterization of a yeast nucleolar protein that is similar to a rat liver nucleolar protein

We have produced monoclonal antibodies against purified nuclei from the yeast Saccharomyces cerevisiae and have characterized three different antibodies that recognize a protein with an apparent molecular weight of 38,000, termed p38. Subcellular fractionation shows that virtually all of p38 occurs in the nuclear fraction. High concentrations of salt (1 M) or urea (6 M) effectively solubilize p38 from a nuclear envelope fraction prepared by digestion of nuclei with DNase. Indirect immunofluorescence demonstrates a crescent shaped distribution of p38 at the inner periphery of the nucleus, with p38 extending between dividing pairs of cells during (closed) mitosis. Postembedding immunogold electron microscopy shows decoration of the densely stained "crescent" region of the yeast nucleus, confirming the localization of p38 to the nucleolus. One of the monoclonals, D77, cross reacts on immunoblots with a single protein of molecular weight 37,000 from purified rat liver nuclei. Indirect immunofluorescence localizes this protein to the nucleolus, and shows that it is dispersed throughout the cell during mitosis. The yeast and rat liver nucleolar proteins behave similarly when electrophoresed in two dimensions, and appear to have basic pI values. Analysis of immunological cross-reactivity using D77, and antibodies specific for nucleolar proteins from other sources, suggests that the rat liver protein is fibrillarin, and demonstrates that p38 shares epitopes with fibrillarin, as well as with other vertebrate nucleolar proteins.     

A karyological study of swamp and dry valley populations of Siberian fir (<Emphasis Type="Italic">Abies sibirica</Emphasis> Ledeb.)

The karyological data on populations of Siberian fir (Abies sibirica Ledeb.) growing in lowland swamp and dry valleys are given. The diploid set of both populations included 24 chromosomes (2n = 24). Polykaryogram analysis revealed seven pairs of metacentric and five pairs of submetacentric chromosomes. The revealed differences between the populations included the absolute length of chromosomes, number of nucleolar organizer regions, and number of nucleoli. Changed chromosome numbers (mixoploidy and aneuploidy) as well as chromosomal aberrations were recorded. For the first time, mitosis was studied in this species and anaphase/telophase aberrations were revealed. The population of Siberian fir growing under extreme conditions of lowland swamp featured the widest range of mutations.Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 1, 2005, pp. 23–29.Original Russian Text Copyright © 2005 by Sedelnikova, Pimenov.     

The relation of DNA synthesis and mitosis in tobacco pith tissue cultured in vitro

Summary DNA synthesis and mitosis were initiated in cultured tobacco pith tissue by means of IAA and kinetin. DNA classes were determined by microspectrophotometric measurements (Feulgen); autoradiographs (tritiated thymidine) served to ascertain whether or not nuclei had undergone DNA synthesis during culture.All mitoses in new cells (resulting from divisions in culture) were diploid and had been preceded by DNA synthesis in culture.Whereas many of the old cells (which had not previously divided in culture) found in diploid or polyploid mitosis had undergone DNA synthesis during culture, others had not. Such non-radioactive mitoses still occurred after 16 days.In view of this, a 4 C nucleus in differentiated tissue should be considered as potentially both diploid and tetraploid, for it appears impossible to predict whether it would, upon restoration of conditions conducive to DNA synthesis and mitosis, enter a diploid mitosis or, after undergoing DNA synthesis, a tetraploid one.A high nuclear DNA content seems to have a much more inhibiting effect on the onset of DNA doubling than on that of mitosis.Somatic polyploidization is understood as the result of two DNA doublings between which mitosis was omitted, or aborted, or in effect undone by a failure of cytokinesis leading to fusion during a later mitosis.This work has been supported by research grants to K. Patau from the U.S. Public Health Service (grant No. C-3313) and the American Cancer Society.     

A limited number of chromosomes makes a nucleus competent to respond to inducers of replication and mitosis in a plant.

Multinucleate tetraploid cells with unbalanced chromosomal distribution in aneuploid nuclei were obtained in Allium cepa L. root meristems. For this, their natural diploid cells were treated with a multipolarizing agent (1 h carbetamide) followed by an inhibitor of cytokinesis (1 h caffeine). Data from these multinucleate cells with aneuploid nuclei suggest that only four out of the thirty-two chromosomes of their autotetraploid complement possess DNA sequences making the nucleus competent to respond to inducers of replication and mitosis. Direct observation of cells where a single replicated chromosome had reached mitosis showed that this chromosome was the one bearing the nucleolar organizer. Six specific chromosomes would confer competence to the nucleus to respond to inducers of replication but not to those producing chromosome condensation. Another four different chromosomes would confer the nucleus with the ability to respond to mitotic inducers but not to replication inducers. The rest of the chromosomal complement seemed to lack any of the DNA sequences needed for these two important cycle transitions. In a nutshell, certain DNA sequences distributed in a few chromosomes of the onion complement are an intranuclear requirement to initiate replication and mitosis in these plant cells.     

Centromere sizes,positions, and movements in the interphase nucleus

Each microspore of the onion Allium fistulosum (n=8) has 8 chromosomes. It is shown that in the microspore the 8 centromeres aggregate to form 2 or 3 centromeric structures. Subsequently, at early mitotic prophase, these aggregates are resolved into 8 separate centromeres and each becomes structurally double. After mitosis the pollen grain contains 2 nuclei, each with 8 separate and distinct centromeres, clustered at the nuclear envelope. As interphase progresses the centromeres of the vegetative nucleus are no longer at the nuclear envelope and they aggregate into 3 or 4 centromeric masses. In the generative nucleus there is less movement. The interphase centromere movements occur in the absence of microtubules. The centromeres range in size from about 0.10 to 0.17 m³ with an average of 0.14 m³ per centromere.     

Electron Microscopic Studies on the Silver-stained Nucleolar Cycle of Physarum polycephalum

The dynamic changes of nucleolar ultrastructure in the cell cycle of Physarum polycephalum Schw. were studied by an en bloc silver-staining method. The results showed that the nucleolus was large in size and situated in the center of the nucleus in late G2-phase, and the fibrillar centers, dense fibrillar components and granular components could be observed in the nucleolus. During prophase, the nucleolus moved towards the periphery of the nucleus and in late prophase disintegrated near the nuclear envelope. In metaphase, the disintegrated nucleolar components were dispersed in masses and located at the periphery of the chromosomal region of the nucleus. No specifically silver-stained area and argentophilic protein sheath were observed on the chromosomes, but there were some big dispersed silver particles within the chromosomes. During telophase the nucleolar components moved towards the two poles along with the chromosomes and co-existed with the decondensing chromatin in daughter nuclei. The nucleolar components then gradually converged with one another and separated from the chromatin. A big nucleolus was formed in the nucleus about 120 min after the completion of mitosis.