Chromosome Disjunction Research Articles

Absence of synapsis during pachynema of the normal sized sex chromosomes of Microtus arvalis.

The pachytene behavior of the chromosomes of males of Microtus arvalis (Pall.) (Rodentia, Arvicolidae) was examined by electron microscopy in microspread preparations of spermatocytes. There was no synapsis between the axes of these two chromosomes during this period. Since synapsis is universally considered a prerequisite for crossing over and chiasmata formation, disjunction of the sex chromosomes in this species prerequisite for crossing over and chiasmata formation, disjunction of the sex chromosomes in this species must be presumed to be achiasmatic. Unlike previously examined species with no synapsis or crossing over between the X and Y, the sex chromosomes of M. arvalis are of normal size: the X chromosome is of an "original" X size and the Y is a small acrocentric. C-band studies of M. arvalis mitotic metaphase reveal no blocks of heterochromatin on the sex chromosomes. The implications of these findings are discussed.

TheCaenorhabditis elegans rsd-2andrsd-6Genes Are Required for Chromosome Functions During Exposure to Unfavorable Environments

In Caenorhabditis elegans, exogenous dsRNA can elicit systemic RNAi, a process that requires the function of many genes. Considering that the activities of many of these genes are also required for normal development, it is surprising that exposure to high concentrations of dsRNA does not elicit adverse consequences to animals. Here, we report inducible phenotypes in attenuated C. elegans strains reared in environments that include nonspecific dsRNA and elevated temperature. Under these conditions, chromosome integrity is compromised in RNAi-defective strains harboring mutations in rsd-2 or rsd-6. Specifically, rsd-2 mutants display defects in transposon silencing, while meiotic chromosome disjunction is affected in rsd-6 mutants. RSD-2 proteins localize to multiple cellular compartments, including the nucleolus and cytoplasmic compartments that, in part, are congruent with calreticulin and HAF-6. We considered that the RNAi defects in rsd-2 mutants might have relevance to membrane-associated functions; however, endomembrane compartmentalization and endocytosis/exocytosis markers in rsd-2 and rsd-6 mutants appear normal. The mutants also possess environmentally sensitive defects in cell-autonomous RNAi elicited from transgene-delivered dsRNAs. Thus, the ultimate functions of rsd-2 and rsd-6 in systemic RNAi are remarkably complex and environmentally responsive.

Chromosome nondisjunction in Drosophila strains mutant for tumor suppressor Merlin

The effect of mutation for gene Merlin on chromosome disjunction in Drosophila during meiosis was genetically studied. Chromosome nondisjunction was not registered in females heterozygous for this mutation and containing structurally normal X chromosomes. In cases when these females additionally contained inversion in one of chromosomes X, a tendency toward the appearance of nondisjunction events was observed in individuals containing mutation in the heterozygote. The genetic construct was obtained allowing the overexpression of protein corresponding to a sterile allele Mer3 in the germ cell line. This construct relieves the lethal effect of Mer4 mutation. The ectopic expression of this mutant protein leads to chromosome nondisjunction in male meiosis.

Condensation of rye chromatin in somatic interphase nuclei of Ph1 and ph1b wheat

The Ph1 locus in hexaploid wheat (Triticum aestivum L.) enforces diploid-like behavior in the first metaphase of meiosis. To test the hypothesis that this chromosome pairing control is exercised by affecting the degree of chromatin condensation, the dispersion of rye chromatin in interphase nuclei in somatic tissues of wheat-rye chromosome translocations 1RS.1BL, 2RS.2BL, 2BS.2RL, 3RS.3DL and 5RS.5BL was compared in Ph1 and ph1b isogenic backgrounds. No significant differences in rye chromatin condensation that could be attributed to the Ph1 locus were detected. Regardless of the Ph1 status, each rye chromosome arm tested conformed to the general Rabl’s orientation and occupied portions of the nuclei proportional to their length. Earlier observations that indicated the involvement of Ph1 locus in rye chromatin condensation in wheat could have been due either to specific loci on the studied 5RL rye arm that control the chromosome condensation process or to damage to the genetic system controlling chromatin condensation in the existing ph1b stocks of wheat. That damage might have been caused by homoeologous recombination and uneven disjunction of chromosomes from multivalents.

Expression and functional analysis of AtMUS81 in Arabidopsis meiosis reveals a role in the second pathway of crossing‐over

Meiotic crossovers/chiasmata, that are required to ensure chromosome disjunction, arise via the class I interference-dependent pathway or via the class II interference-free pathway. The proportions of these two classes vary considerably between different organisms. In Arabidopsis, about 85% of chiasmata are eliminated in Atmsh4 mutants, denoting that these are class I events. In budding and fission yeasts Msh4-independent crossovers arise largely or entirely via a Mus81-dependent pathway. To investigate the origins of the 15% residual (AtMSH4-independent) chiasmata in Arabidopsis we conducted a cytological and molecular analysis of AtMUS81 meiotic expression and function. Although AtMUS81 functions in somatic DNA repair and recombination, it is more highly expressed in reproductive tissues. The protein is abundantly present in early prophase I meiocytes, where it co-localizes, in a double-strand break-dependent manner, with the recombination protein AtRAD51. Despite this, an Atmus81 mutant shows normal growth and has no obvious defects in reproductive development that would indicate meiotic impairment. A cytological analysis confirmed that meiosis was apparently normal in this mutant and its mean chiasma frequency was similar to that of wild-type plants. However, an Atmsh4/Atmus81 double mutant revealed a significantly reduced mean chiasma frequency (0.85 per cell), compared with an Atmsh4 single mutant (1.25 per cell), from which we conclude that AtMUS81 accounts for some, but not all, of the 15% AtMSH4-independent residual crossovers. It is possible that other genes are responsible for these residual chiasmata. Alternatively the AtMUS81 pathway coexists with an alternative parallel pathway that can perform the same functions.

Analysis of Intra-specific Genetic Variation in Calligonum polygonoides L. (Polygonaceae)-A Keystone Species of Indian Desert

Calligonum polygonoides is an endemic and threatened species reported from semi-arid and arid zones of Rajasthan and adjoining desert belt especially on loose sand dunes. Polygonaceae, the family to which C. polygonoides belongs has about 12 genera distributed in India which comprises about 115 species, distributed chiefly in north temperate, arctic and southern regions, whereas a few are spread over tropical regions. This is the first attempt to determine the gametic chromosome number of C. polygonoides from various populations of arid zone regions in western Rajasthan. The genetic variation analyzed is based on chromosome associations, chiasma frequency at diplotene/diakinesis/metaphase I and pattern of disjunction of chromosomes at anaphase I.

Meiotic pairing and disjunction of mini-X chromosomes in drosophila is mediated by 240-bp rDNA repeats and the homolog conjunction proteins SNM and MNM.

In most eukaryotes, segregation of homologous chromosomes during meiosis is dependent on crossovers that occur while the homologs are intimately paired during early prophase. Crossovers generate homolog connectors known as chiasmata that are stabilized by cohesion between sister-chromatid arms. In Drosophila males, homologs pair and segregate without recombining or forming chiasmata. Stable pairing of homologs is dependent on two proteins, SNM and MNM, that associate with chromosomes throughout meiosis I until their removal at anaphase I. SNM and MNM localize to the rDNA region of the X-Y pair, which contains 240-bp repeats that have previously been shown to function as cis-acting chromosome pairing/segregation sites. Here we show that heterochromatic mini-X chromosomes lacking native rDNA but carrying transgenic 240-bp repeat arrays segregate preferentially from full-length sex chromosomes and from each other. Mini-X pairs do not form autonomous bivalents but do associate at high frequency with the X-Y bivalent to form trivalents and quadrivalents. Both disjunction of mini-X pairs and multivalent formation are dependent on the presence of SNM and MNM. These results imply that 240-bp repeats function to mediate association of sex chromosomes with SNM and MNM.

Cyclin-B1-mediated inhibition of excess separase is required for timely chromosome disjunction

Separase, the cysteine protease that cleaves cohesin and thereby triggers chromosome disjunction, is inhibited by both securin- and phosphorylation-dependent cyclin B1 binding. Using a novel phosphorylation-specific antibody, we show that mitotic-specific phosphorylation of human separase on S1126 is required to establish, but not maintain, cyclin B1 binding. Cells expressing a non-phosphorylatable S1126A mutant maintain cohesion early in mitosis, aligning their chromosomes. Cohesion is then synchronously lost 5 minutes ahead of schedule, without degrading securin or cyclin B1. This premature chromatid disjunction requires the catalytic activity of separase, indicating that it is dependent on cohesin cleavage. Single chromatids then attempt to realign but the lack of tension results in unstable kinetochore-microtubule interactions and Aurora-B-dependent spindle checkpoint activation. Separase mutants that cannot bind cyclin B1 but are phosphorylated on S1126 phenocopy separase S1126A, indicating that cyclin B1 binding, rather than phosphorylation, is the key inhibitory event. Significantly, by overexpressing separase S1126A, we have simultaneously overridden the two known inhibitory mechanisms. First, by elevating separase levels above securin, securin-mediated inhibition is alleviated. Second, by preventing phosphorylation, cyclin-B1-mediated inhibition is also alleviated. Surprisingly, however, cohesion is maintained during the early stages of mitosis, indicating the existence of another mechanism that either inhibits separase or protects its substrate during early mitosis.

Cytogenetics of species of Chamaecrista (Leguminosae – Caesalpinioideae) native to southern Brazil

Chromosome numbers, karyotypes, meiotic behaviour and pollen analysis are presented for species of Chamaecrista Moench (Leguminosae, Caesalpinioideae, Cassieae) native to southern Brazil: C. nictitans ssp. patellaria, C. nictitans ssp. disadena, C. repens, C. rotundifolia, C. flexuosa, C. vestita and C. desvauxii. Meiotic behaviour is reported for the first time for all the taxa and was very regular; only bivalents were formed at diakinesis and metaphase I, chromosome disjunction and segregation were regular at anaphases I and II, meiotic indexes were over 99% and pollen fertility was over 92%. Pollen grains were subprolate in C. flexuosa and C. vestita and prolate–spheroidal in the other taxa. Karyotypes were symmetrical in all six species and the data are original, except for C. nictitans ssp. patellaria. Chromosome number is presented for the first time for C. repens (2n = 16) and has been confirmed for the other taxa: 2n = 14 for C. desvauxii, 2n = 32 for the tetraploid C. nictitans ssp. patellaria and C. nictitans ssp. disadena, and 2n = 16 for the other species. These two basic numbers found in the genus, x = 7 and x = 8, point to chromosome evolution by dysploidy, which has also been accompanied by polyploidy. © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society, 2006, 150, 429–439.

Frequency of 677C -> T and 1298A -> C polymorphisms in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene in Turner syndrome individuals

Turner syndrome (TS) is an interesting model for investigating the association between methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms and non-disjunction because of the high frequency of chromosomal mosaicism among patients with this syndrome. We determined the frequencies of MTHFR 677C -> T and 1298A -> C polymorphic mutations in 49 patients with TS and 200 control individuals. The frequency of the 677C -> T allele was 0.39 for patients and 0.29 for controls while that of the 1298A -> C allele was 0.28 for patients and 0.25 for controls. Genotype frequencies were shown to be different in patients and controls (chi2 = 12.143; p = 0.033), and this was attributable to the higher frequency of the C677C -> T /677C -> T genotype among TS patients. In homozygotes, this mutation might have an effect on somatic chromosome disjunction by decreasing MTHFR activity.

Differential Regulation of Strand-Specific Transcripts from Arabidopsis Centromeric Satellite Repeats

Centromeres interact with the spindle apparatus to enable chromosome disjunction and typically contain thousands of tandemly arranged satellite repeats interspersed with retrotransposons. While their role has been obscure, centromeric repeats are epigenetically modified and centromere specification has a strong epigenetic component. In the yeast Schizosaccharomyces pombe, long heterochromatic repeats are transcribed and contribute to centromere function via RNA interference (RNAi). In the higher plant Arabidopsis thaliana, as in mammalian cells, centromeric satellite repeats are short (180 base pairs), are found in thousands of tandem copies, and are methylated. We have found transcripts from both strands of canonical, bulk Arabidopsis repeats. At least one subfamily of 180–base pair repeats is transcribed from only one strand and regulated by RNAi and histone modification. A second subfamily of repeats is also silenced, but silencing is lost on both strands in mutants in the CpG DNA methyltransferase MET1, the histone deacetylase HDA6/SIL1, or the chromatin remodeling ATPase DDM1. This regulation is due to transcription from Athila2 retrotransposons, which integrate in both orientations relative to the repeats, and differs between strains of Arabidopsis. Silencing lost in met1 or hda6 is reestablished in backcrosses to wild-type, but silencing lost in RNAi mutants and ddm1 is not. Twenty-four–nucleotide small interfering RNAs from centromeric repeats are retained in met1 and hda6, but not in ddm1, and may have a role in this epigenetic inheritance. Histone H3 lysine-9 dimethylation is associated with both classes of repeats. We propose roles for transcribed repeats in the epigenetic inheritance and evolution of centromeres.

Chromosome Morphogenesis: Condensin-Dependent Cohesin Removal during Meiosis

During meiosis, segregation of homologous chromosomes necessitates the coordination of sister chromatid cohesion, chromosome condensation, and recombination. Cohesion and condensation require the SMC complexes, cohesin and condensin, respectively. Here we use budding yeast Saccharomyces cerevisiae to show that condensin and Cdc5, a Polo-like kinase, facilitate the removal of cohesin from chromosomes prior to the onset of anaphase I when homologs segregate. This cohesin removal is critical for homolog segregation because it helps dissolve the recombination-dependent links between homologs that form during prophase I. Condensin enhances the association of Cdc5 with chromosomes and its phosphorylation of cohesin, which in turn likely stimulates cohesin removal. Condensin/Cdc5-dependent removal of cohesin underscores the potential importance of crosstalk between chromosome structural components in chromosome morphogenesis and provides a mechanism to couple chromosome morphogenesis with other meiotic events.

Hydrostatic pressure treatment during the first mitosis does not suppress the first cleavage but the second one

Suppression of cell division causes chromosome set doubling. Some chemical agents or physical shocks such as temperature or hydrostatic pressure are effective tools for suppression of cell division. As spindles are obviously inactivated or disorganized by these treatments, it has been supposed that inactivation or disassembly of spindles blocks the anaphase movement of chromosomes and a duplicated nucleus is formed without cell division. The present study demonstrated that hydrostatic pressure treatment (650 kg/cm 2 for 6 min) around the time of metaphase of the first cell cycle of the rainbow trout embryos did not suppress the first cleavage but the second one. Spindles disassembled by the hydrostatic pressure or heat shock regenerated soon after treatment, resulting in the occurrence of the first mitosis. Interestingly, a monopolar spindle was assembled in each blastomere in the second cell cycle, and disjunction of duplicated chromosomes and the cleavage was prevented, leading to the chromosome set doubling. From the third cell cycle, normal cell division resumed. No significant difference was found between the area of the nucleus plate of the treated embryos and twice the area of the nucleus plate of control embryos in the third cell cycle, meaning that the chromosome sets had been doubled at the end of the second cell cycle. The process of chromosome set doubling caused by heat shock seemed to be fundamentally the same as that caused by hydrostatic pressure. To the best of our knowledge, this is the first time that the mechanism of chromosome set doubling in animal eggs treated with hydrostatic-pressure or heat shock has been clarified. Haploid–diploid or diploid–tetraploid mosaics sometimes occur among individuals treated for cleavage inhibition. The mechanism of such occurrence of mosaicism is, however, not clear. In this study, we found interesting two-cell stage embryos, which had a monopolar spindle in one blastomere and a bipolar spindle in the other during the second mitosis in a batch subjected to tetraploidization treatment. These embryos have a high potential of developing diploid–tetraploid mosaics. This paper also discusses the mechanism of occurrence of these aberrant embryos and discusses their relationship to diploid–tetraploid mosaicism.

Isolation of a Tetrahymena thermophila strain which induced metaphase I meiotic arrest: new pathway of abortive conjugation.

A hypodiploid strain of Tetrahymena thermophila has been obtained that shows arrest at the stage of condensed nuclei, corresponding to metaphase I of normal conjugants and induced arrest at meiotic metaphase I (i.e. at the stage of condensed, bivalent chromosomes) in its wt partner mate. The metaphase I arrested conjugants retained their old macronuclei and most of them underwent cell fusion, instead of separation of exconjugants. The doublets were viable and cortically integrated. When the arrest inducing strain was crossed to the haploid tester strain, the haploid micronuclei were arrested in the meiotic metaphase I as the diploid ones had been; the monovalent, chromosomes were condensed, the arms of sister chromatids were not separated, and they were not segregated. Separation of the arms of sister chromatids and disjunction of bivalent chromosomes were not prerequisite for the formation of microtubular spindles in those cells that were arrested in meiotic metaphase I. After re-feeding, the doublet cells resumed cell divisions, segregating two macronuclei and micronuclei at random. One macronucleus was derived from the arrest inducing strain and the other from the tester strain. Heterokaryon strains with macronuclei derived from the parental arrest inducing strain and with the micronucleus derived from the parental wt tester strain were obtained. Surprisingly, these heterokaryons did not induce meiotic arrest. Thus, the arrest in the melotic metaphase I was induced by the micronucleus and not by the macronucleus of the arrest inducing strain.

Three classes of genes mutated in colorectal cancers with chromosomal instability.

Although most colorectal cancers are chromosomally unstable, the basis for this instability has not been defined. To determine whether genes shown to cause chromosomal instability in model systems were mutated in colorectal cancers, we identified their human homologues and determined their sequence in a panel of colorectal cancers. We found 19 somatic mutations in five genes representing three distinct instability pathways. Seven mutations were found in MRE11, whose product is involved in double-strand break repair. Four mutations were found among hZw10, hZwilch/FLJ10036, and hRod/KNTC, whose products bind to one another in a complex that localizes to kinetochores and controls chromosome segregation. Eight mutations were found in Ding, a previously uncharacterized gene with sequence similarity to the Saccharomyces cerevisiae Pds1, whose product is essential for proper chromosome disjunction. This analysis buttresses the evidence that chromosomal instability has a genetic basis and provides clues to the mechanistic basis of instability in cancers.

Anaphase-promoting complex in Caenorhabditis elegans.

Anaphase-promoting complex in Caenorhabditis elegans.

Gamma-tubulin plays an essential role in the coordination of mitotic events.

Recent data from multiple organisms indicate that gamma-tubulin has essential, but incompletely defined, functions in addition to nucleating microtubule assembly. To investigate these functions, we examined the phenotype of mipAD159, a cold-sensitive allele of the gamma-tubulin gene of Aspergillus nidulans. Immunofluorescence microscopy of synchronized material revealed that at a restrictive temperature mipAD159 does not inhibit mitotic spindle formation. Anaphase A was inhibited in many nuclei, however, and after a slight delay in mitosis (approximately 6% of the cell cycle period), most nuclei reentered interphase without dividing. In vivo observations of chromosomes at a restrictive temperature revealed that mipAD159 caused a failure of the coordination of late mitotic events (anaphase A, anaphase B, and chromosomal disjunction) and nuclei reentered interphase quickly even though mitosis was not completed successfully. Time-lapse microscopy also revealed that transient mitotic spindle abnormalities, in particular bent spindles, were more prevalent in mipAD159 strains than in controls. In experiments in which microtubules were depolymerized with benomyl, mipAD159 nuclei exited mitosis significantly more quickly (as judged by chromosomal condensation) than nuclei in a control strain. These data reveal that gamma-tubulin has an essential role in the coordination of late mitotic events, and a microtubule-independent function in mitotic checkpoint control.

Nucleotide excision repair deficiency causes elevated levels of chromosome gain in Saccharomyces cerevisiae

Aneuploidy is the most frequent aberration observed in tumor cells, and underlies many debilitating and cancer-prone congenital disorders. Aneuploidy most often arises as a consequence of chromosomal non-disjunction, however, little is known about the genetic and epigenetic factors that affect the chromosomal segregation process. As many cancer-prone syndromes are associated with defects in DNA repair pathways we decided to investigate the relationship between DNA repair in mutation avoidance pathways, namely base and nucleotide excision, and mismatch repair (MMR), and aneuploidy in the yeast Saccharomyces cerevisiae. Isogenic haploid and diploid DNA repair deficient yeast strains were constructed, and spontaneous levels of intra- and inter-chromosomal recombination, forward mutation, chromosome gain, and loss were measured. We show that the nucleotide excision repair (NER) pathway is required for accurate chromosomal disjunction. In the absence of Rad1, Rad2, or Rad4, spontaneous levels of chromosome XV gain were significantly elevated in both haploid and diploid mutant strains. Thus, chromosome gain may be an additional cancer predisposing event in NER deficient patients.

A deficiency screen of the major autosomes identifies a gene (matrimony) that is haplo-insufficient for achiasmate segregation in Drosophila oocytes.

In Drosophila oocytes, euchromatic homolog-homolog associations are released at the end of pachytene, while heterochromatic pairings persist until metaphase I. A screen of 123 autosomal deficiencies for dominant effects on achiasmate chromosome segregation has identified a single gene that is haplo-insufficient for homologous achiasmate segregation and whose product may be required for the maintenance of such heterochromatic pairings. Of the deficiencies tested, only one exhibited a strong dominant effect on achiasmate segregation, inducing both X and fourth chromosome nondisjunction in FM7/X females. Five overlapping deficiencies showed a similar dominant effect on achiasmate chromosome disjunction and mapped the haplo-insufficient meiotic gene to a small interval within 66C7-12. A P-element insertion mutation in this interval exhibits a similar dominant effect on achiasmate segregation, inducing both high levels of X and fourth chromosome nondisjunction in FM7/X females and high levels of fourth chromosome nondisjunction in X/X females. The insertion site for this P element lies immediately upstream of CG18543, and germline expression of a UAS-CG18543 cDNA construct driven by nanos-GAL4 fully rescues the dominant meiotic defect. We conclude that CG18543 is the haplo-insufficient gene and have renamed this gene matrimony (mtrm). Cytological studies of prometaphase and metaphase I in mtrm hemizygotes demonstrate that achiasmate chromosomes are not properly positioned with respect to their homolog on the meiotic spindle. One possible, albeit speculative, interpretation of these data is that the presence of only a single copy of mtrm disrupts the function of whatever "glue" holds heterochromatically paired homologs together from the end of pachytene until metaphase I.

Crossover Interference in Humans

Crossing-over between homologous chromosomes facilitates proper disjunction of chromosomes during meiosis I. In many organisms, gene functions that are essential to crossing-over also facilitate the intimate chromosome pairing called "synapsis." Many organisms--including budding yeast, humans, zebrafish, Drosophila, and Arabidopsis--regulate the distribution of crossovers, so that, most of the time, each chromosome bundle gets at least one crossover while the mean number of crossovers per chromosome remains modest. This regulation is obtained through crossover interference. Recent evidence suggests that the organisms that use recombination functions to achieve synapsis have two classes of crossovers, only one of which is subject to interference. We statistically test this two-pathway hypothesis in the CEPH data and find evidence to support the two-pathway hypothesis in humans.