Chromosomes In Meiosis Research Articles

Components of the RNAi Machinery That Mediate Long-Distance Chromosomal Associations Are Dispensable for Meiotic and Early Somatic Homolog Pairing inDrosophila melanogaster

Homolog pairing is indispensable for the proper segregation of chromosomes in meiosis but the mechanism by which homologs uniquely pair with each other is poorly understood. In Drosophila, somatic chromosomes also undergo full homolog pairing by an unknown mechanism. It has been recently demonstrated that both insulator function and somatic long-distance interactions between Polycomb response elements (PREs) are stabilized by the RNAi machinery in Drosophila. This suggests the possibility that long-distance pairing interactions between homologs, either during meiosis or in the soma, may be stabilized by a similar mechanism. To test this hypothesis, we have characterized meiotic and early somatic chromosome pairing of homologous chromosomes in flies that are mutant for various components of the RNAi machinery. Despite the identification of a novel role for the piRNA machinery in meiotic progression and synaptonemal complex (SC) assembly, we have found that the components of the RNAi machinery that mediate long-distance chromosomal interactions are dispensable for homologous chromosome pairing. Thus, there appears to be at least two mechanisms that bring homologous sequences together within the nucleus: those that act between dispersed homologous sequences and those that act to align and pair homologous chromosomes.

Chromosome Mechanics and Meiotic Engine Maintenance

Chromosome Mechanics and Meiotic Engine Maintenance

Features of the genetic structure of the earthworms Aporrectodea (Superspecies) Caliginosa (Oligochaeta: Lumbricidae) complex in Ukraine

An investigation of the genetic structure of the Aporrectodea caliginosa-A. trapezoids diploidpolyploid complex of earthworms found in the Ukraine is performed by means of biochemical genetic marking with respect to six loci (Aat, Es-1,-2,-3, and-4, and Mdh) and karyotyping. All 646 individuals from 21 samples are analyzed at the biochemical gene level and karyotype samples are obtained from 70 specimens. As a result, diploid amphimictic A. caliginosa individuals (2n = 36), which form panmictic populations, and triploid (2n = 54) A. trapezoides individuals, represented by 19 hypothetical clones discovered in the course of an analysis of 157 specimens, are clearly differentiated. A clear trend towards dominance of the triploid forms in the steppe zone of Ukraine is discovered. Here they represent roughly 70% as against 12% of all A. (superspecies) caliginosa individuals in the northern forest regions. Based on the stated nature of the heterozygosity of the loci studied and the behavior of the chromosomes in meiosis, it may be claimed that the cloned forms are allotriploids, formed as a result of hybridization of a series of related forms, which, judging from the allelic pools, does not include amphimictic species that now inhabit the territory of Ukraine. That is, either the apomictic clones of A. trapezoides lack a local origin and their appearance among the fauna of Ukraine is a consequence of settlement in arid steppe regions that are unfavorable for earthworms or, on the other hand, amphimictic A. caliginosa is an invasive species.

The Kleisin Subunit of Cohesin Dictates Damage-Induced Cohesion

Cohesin, the protein complex that mediates sister chromatid cohesion, is required for faithful chromosome segregation and efficient repair of double-strand breaks (DSBs). Cohesion generation is normally restricted to S phase. However, in G2/M, a DSB activates cohesion generation near the DSB and genome-wide. Here, using budding yeast, we show that DSB-induced cohesion occurs when cohesin contains the kleisin subunit, Mcd1 (Scc1), but not when Mcd1 is replaced by its meiotic isoform, Rec8. We exploit this divergence to demonstrate that serine 83 of Mcd1 and the Chk1 kinase are critical determinants for DSB-induced cohesion. We propose that a DSB in G2/M activates Mec1 (ATR), which in turn stimulates Chk1-dependent phosphorylation of Mcd1 at serine 83. Serine 83 phosphorylation promotes chromatin-bound cohesin to become cohesive.

Orientation of Ts65Dn Chromosome in Meiosis I Predicts Reduction of Metaphase II Spermatocytes in a Mouse Model of Down Syndrome.

Orientation of Ts65Dn Chromosome in Meiosis I Predicts Reduction of Metaphase II Spermatocytes in a Mouse Model of Down Syndrome.

Genetic regulation of the centromere division in rye and wheat univalent chromosomes in dimonosomics during the meiotic anaphase I

Cytogenetic analysis of meiosis in the wheat--rye dimonosomics 1Rv-1A, 1Ron-1A, 2R-2D, 5R-5A, and 6R-6A was conducted. C-banding was used to study the segregation pattern of each of two univalent chromosomes during the first meiotic division. It has been shown that the division frequency of the centromeric regions of all rye chromosomes in the pair studied is significantly higher than in the wheat chromosomes. The ANOVA performed suggest that the plant genotype contributes significantly (at P = 0.05) to the behavior pattern of univalent chromosomes in meiosis. The data obtained demonstrate that the rye and wheat chromosomes studied are involved in genetic regulation of centromere division in meiotic anaphase I (AI). The presence of rye chromosome 2R and wheat chromosome 2D suppresses the division of centromeres of the sister chromatids in AI. Rye chromosomes 1Rv, 1Ron, 5R, and 6R induce equational division; however, rye chromosome 1Rv increases to a greater degree the frequency of equational division of wheat chromosome 1A as compared with chromosome 1Ron.

The B chromosome in the meiotic process of the African pest grasshopper <i>Taphronota thaelephora</i> STAL. (Orthoptera: Pyrgomorphidae).

This paper presents the first record of B chromosomes in some adult male individuals of the African pest grasshopper Taphronota thaelephora collected from Balepa – Mbouda in the Western Province of Cameroon. Of the twenty individuals examined, thirteen had the characteristic Pyrgomorphidae karyotype of 2N= 19(18A+XO) while the remaining seven had a diploid chromosome number of 2N= 20(18A+XO+1B) indicating the presence of a single B chromosome. The B chromosome in this population was short, acrocentric in morphology and stained to the same intensity as the autosomes throughout meiosis. It did not associate with any chromosome in meiosis. In some cells it migrated to the same pole as the X-chromosome while in other cells it migrated to the pole opposite to that of the X-chromosome indicating irregular behaviour in meiosis. The presence of the B chromosome did not affect chiasma frequency in the species. International Journal of Biological & Chemical Sciences Vol. 1 (2) 2007: pp. 151-157

Rescuing distal crossovers

Optimally positioned crossovers facilitate the proper segregation of chromosomes in meiosis. A new study in yeast implicates the spindle checkpoint in the rescue of crossovers that occur too distant from the centromere, possibly shedding light on the origins of nondisjunction in higher organisms.

Bi-orientation of achiasmatic chromosomes in meiosis I oocytes contributes to aneuploidy in mice

The spindle assembly checkpoint guards against chromosomal missegregation but does not induce arrest in oocytes that contain a few achiasmatic chromosomes (univalents). We followed the fate of univalents in oocytes during the first meiotic division, and although these preserved a meiotic kinetochore structure, they were also bi-oriented in a mitotic manner. The hybrid chromosomal configuration attained by univalents allows them to evade the spindle assembly checkpoint and contribute to aneuploidy in oocytes.

Centromeres were derived from telomeres during the evolution of the eukaryotic chromosome.

The centromere is the DNA region of the eukaryotic chromosome that determines kinetochore formation and sister chromatid cohesion. Centromeres interact with spindle microtubules to ensure the segregation of chromatids during mitosis and of homologous chromosomes in meiosis. The origin of centromeres, therefore, is inseparable from the evolution of cytoskeletal components that distribute chromosomes to offspring cells. Although the origin of the nucleus has been debated, no explanation for the evolutionary appearance of centromeres is available. We propose an evolutionary scenario: The centromeres originated from telomeres. The breakage of the ancestral circular genophore activated the transposition of retroelements at DNA ends that allowed the formation of telomeres by a recombination-dependent replication mechanism. Afterward, the modification of the tubulin-based cytoskeleton that allowed specific subtelomeric repeats to be recognized as new cargo gave rise to the first centromere. This switch from actin-based genophore partition to a tubulin-based mechanism generated a transition period during which both types of cytoskeleton contributed to fidelity of chromosome segregation. During the transition, pseudodicentric chromosomes increased the tendency toward chromosomal breakage and instability. This instability generated multiple telocentric chromosomes that eventually evolved into metacentric or holocentric chromosomes.

Reconstruction of the kinetochore: a prelude to meiosis

In eukaryotic organisms, chromosomes are spatially organized within the nucleus. Such nuclear architecture provides a physical framework for the genetic activities of chromosomes, and changes its functional organization as the cell moves through the phases of the cell cycle. The fission yeast Schizosaccharomyces pombe provides a striking example of nuclear reorganization during the transition from mitosis to meiosis. In this organism, centromeres remain clustered at the spindle-pole body (SPB; a centrosome-equivalent structure in fungi) during mitotic interphase. In contrast, during meiotic prophase, centromeres dissociate from the SPB and telomeres cluster to the SPB. Recent studies revealed that this repositioning of chromosomes is regulated by mating pheromone signaling. Some centromere proteins disappear from the centromere in response to mating pheromone, leading to dissociation of centromeres from the SPB. Interestingly, mating pheromone signaling is also required for monopolar orientation of the kinetochore which is crucial for proper segregation of sister chromatids during meiosis. When meiosis is induced in the absence of mating pheromone signaling, aberrant chromosome behaviors are observed: the centromere proteins remain at the centromere; the centromere remains associated with the SPB; and sister chromatids segregate precociously in the first meiotic division. These aberrant chromosome behaviors are all normalized by activating the mating pheromone signaling pathway. Thus, action of mating pheromone on the centromere is important for coherent behavior of chromosomes in meiosis. Here we discuss repositioning and reconstruction of the centromere during the transition from mitosis to meiosis, and highlight its significance for proper progression of meiosis.

Differential timing of S phases, X chromosome replication, and meiotic prophase in the C. elegans germ line

The replication of chromosomes in meiosis is an important first step for subsequent chromosomal interactions that promote accurate disjunction in the first of two segregation events to generate haploid gametes. We have developed an assay to monitor DNA replication in vivo in mitotic and meiotic germline nuclei of the nematode Caenorhabditis elegans. Using mutants that affect the mitosis/meiosis switch, we show that meiotic S phase is at least twice as long as mitotic S phase in C. elegans germ cell nuclei. Furthermore, our assay reveals that different regions of the genome replicate at different times, with the heterochromatic-like X chromosomes replicating at a distinct time from the autosomes. Finally, we have exploited S-phase labeling to monitor the timing of progression through meiotic prophase. Meiotic prophase for oocyte production in hermaphrodites lasts 54–60 h. Further, we find that the duration of the pachytene sub-stage is modulated by the presence of sperm. On the other hand, meiotic prophase for sperm production in males is completed by 20–24 h. Possible sources for the sex-specific differences in meiotic prophase kinetics are discussed.

Sequential Loading of Cohesin Subunits during the First Meiotic Prophase of Grasshoppers

The cohesin complexes play a key role in chromosome segregation during both mitosis and meiosis. They establish sister chromatid cohesion between duplicating DNA molecules during S-phase, but they also have an important role during postreplicative double-strand break repair in mitosis, as well as during recombination between homologous chromosomes in meiosis. An additional function in meiosis is related to the sister kinetochore cohesion, so they can be pulled by microtubules to the same pole at anaphase I. Data about the dynamics of cohesin subunits during meiosis are scarce; therefore, it is of great interest to characterize how the formation of the cohesin complexes is achieved in order to understand the roles of the different subunits within them. We have investigated the spatio-temporal distribution of three different cohesin subunits in prophase I grasshopper spermatocytes. We found that structural maintenance of chromosome protein 3 (SMC3) appears as early as preleptotene, and its localization resembles the location of the unsynapsed axial elements, whereas radiation-sensitive mutant 21 (RAD21) (sister chromatid cohesion protein 1, SCC1) and stromal antigen protein 1 (SA1) (sister chromatid cohesion protein 3, SCC3) are not visualized until zygotene, since they are located in the synapsed regions of the bivalents. During pachytene, the distribution of the three cohesin subunits is very similar and all appear along the trajectories of the lateral elements of the autosomal synaptonemal complexes. However, whereas SMC3 also appears over the single and unsynapsed X chromosome, RAD21 and SA1 do not. We conclude that the loading of SMC3 and the non-SMC subunits, RAD21 and SA1, occurs in different steps throughout prophase I grasshopper meiosis. These results strongly suggest the participation of SMC3 in the initial cohesin axis formation as early as preleptotene, thus contributing to sister chromatid cohesion, with a later association of both RAD21 and SA1 subunits at zygotene to reinforce and stabilize the bivalent structure. Therefore, we speculate that more than one cohesin complex participates in the sister chromatid cohesion at prophase I.

Holocentric chromosomes in meiosis. II. The modes of orientation and segregation of a trivalent

The modes of orientation and segregation of the sex chromosome trivalent X1X2Y in male meiosis of Cacopsylla mali (Psylloidea, Homoptera) were analysed. Males with an X1X2Y sex chromosome system coexist with males displaying a neo-XY system in populations of this species. The fusion chromosome resulting in the formation of a trivalent in meiosis originates from the fusion of an autosome with the neo-Y chromosome. In the majority of metaphase I cells (92.4%) the X1X2Y trivalent showed co-orientation; X1 and X2 chromosomes oriented towards one pole whereas the Y oriented towards the opposite pole. In the rest of the cells (7.6%) the trivalent with subterminal chiasmata was oriented parallel to the equatorial plane. From this orientation the trivalent produced triple chromatids joined together by undivided telomeric parts of chromosomes and hence by sister chromatid cohesion at anaphase I. In the majority of metaphase II cells the orientation of triple chromatids suggested the production of unbalanced gametes. However, in a small number of cells (1.7%) the trivalent showed co-orientation of X1X2 with Y. Both the first division and second division co-orientations, or 94.1% of divisions as a whole, were estimated to yield balanced gametes, containing either X1 and X2 chromosomes or Y chromosome. It was concluded that, since the triple chromatid contained undivided telomere regions at metaphase II, which divided at anaphase II, the orientation of the trivalent with its longitudinal axis parallel to the equatorial plane in metaphase I also represents co-orientation and results in pre-reduction. The existence of post-reductional behaviour of holocentric bivalents and multivalents is discussed.

Molecular Activities of Meiosis-specific Proteins Hop2, Mnd1, and the Hop2-Mnd1 Complex

The mouse Hop2 and Mnd1 proteins, which can form a stable heterodimeric complex, ensure the proper synapsis of homologous chromosomes in meiosis by acting in concert with Rad51 and Dmc1 to promote the strand invasion (D-loop formation) step of homologous recombination. Hop2 alone promotes D-loop formation, but Mnd1 and the Hop2-Mnd1 complex do not. Here we show that only the heterodimer complex, but not the individual proteins, can stimulate strand invasion by Dmc1. Furthermore, we demonstrate that the interaction with Mnd1 provokes changes in Hop2 that are responsible not only for abrogating the recombinase activity of Hop2 but also for generating a new molecular interface able to physically interact with and stimulate Dmc1. We also show that coiled-coil motifs in Hop2 and Mnd1 are essential for their interaction with each other and that a clearly delineated region near the COOH terminus of both proteins is necessary for both the DNA binding and single-strand annealing by the Hop-Mnd1 heterodimer. Finally, we describe a point mutation in Hop2 that dissociates its strand invasion activity from its ability to bind and anneal DNA.

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.

Ionizing radiation and genetic risks: XIV. Potential research directions in the post-genome era based on knowledge of repair of radiation-induced DNA double-strand breaks in mammalian somatic cells and the origin of deletions associated with human genomic disorders

Recent estimates of genetic risks from exposure of human populations to ionizing radiation are those presented in the 2001 report of the United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR). These estimates incorporate two important concepts, namely, the following: (1) most radiation-induced mutations are DNA deletions, often encompassing multiple genes, but only a small proportion of the induced deletions is compatible with offspring viability; and (2) the viability-compatible deletions induced in germ cells are more likely to manifest themselves as multi-system developmental anomalies rather than as single gene disorders. This paper: (a) pursues these concepts further in the light of knowledge of mechanisms of origin of deletions and other rearrangements from two fields of contemporary research: repair of radiation-induced DNA double-strand breaks (DSBs) in mammalian somatic cells and human molecular genetics; and (b) extends them to deletions induced in the germ cell stages of importance for radiation risk estimation, namely, stem cell spermatogonia in males and oocytes in females. DSB repair studies in somatic cells have elucidated the roles of two mechanistically distinct pathways, namely, homologous recombination repair (HRR) that utilizes extensive sequence homology and non-homologous end-joining (NHEJ) that requires little or no homology at the junctions. A third process, single-strand annealing (SSA), which utilizes short direct repeat sequences, is considered a variant of HRR. HRR is most efficient in late S and G 2 phases of the cell cycle and is a high fidelity mechanism. NHEJ operates in all cell cycle phases, but is especially important in G 1. In the context of radiation-induced DSBs, NHEJ is error-prone. SSA is also an error-prone mechanism and its role is presumably similar to that of HRR. Studies in human molecular genetics have demonstrated that the occurrence of large deletions, duplications or other rearrangements in certain regions of the genome is related to the presence of large segments of repetitive DNA called segmental duplications (also called duplicons or low copy repeats, LCRs) in such regions. The mechanism that is envisaged for the origin of deletions and other rearrangements involves misalignment of region-specific LCRs of homologous chromosomes in meiosis followed by unequal crossing-over (i.e., non-allelic homologous recombination, NAHR). We hypothesize that: (a) in spermatogonial stem cells, NHEJ is probably the principal mechanism underlying the origin of radiation-induced deletions, although SSA and NAHR may also be involved to some extent, especially at low doses; and (b) in irradiated oocytes, NAHR is likely to be the main mechanism for generating deletions. We suggest future research possibilities, including the development of models for identifying regions of the genome that are susceptible to radiation-induced deletions. Such efforts may have particular significance in the context of the estimation of genetic risks of radiation exposure of human females, a problem that is still with us.

Meiotic observations on the pollen mother cells of Manglietia patungensis, an endangered species

A comparative study of meiosis of pollen mother cells in Manglietia patungensis Hu and its closely related species M. yuyuanensis Law was carried out. These two species had the same chromosome number of 2n=38 and the same karyotype, but their chromosomal behaviors in meiosis were different. In M. patungensis, the typical chromosome configurations observed at metaphase were 2n=0.30IV+18.33II+0.15I, whereas those in M. yuyuanensis were 2n=19II, indicating that some chromosomes in M. patungensis might have undergone inversion and that some chromosome components were possibly paracentric inversion heterozygotes. The abnormal rate of chromosomal behaviors at anaphase I and II was different in the two species. At anaphase II, 8.8% cells were observed with lagging chromosomes which in each cell did not exceed two in M. yuyuanensis, but in M. patungensis, 29.2% cells were observed with lagging chromosomes which could reach 11 in each cell. Additionally, broken chromosomes were observed in M. patungensis. The above results indicate that the abnormal rate of chromosomal behaviors in the meiosis of M. patungensis was higher than that in M. yuyuanensis. The abnormal behaviors of the chromosomes in meiosis observed in M. patungensis may have affected the development of pollen grains in this species.

Meiotic chromosome segregation mutants identified by insertional mutagenesis of fission yeast Schizosaccharomyces pombe; tandem-repeat, single-site integrations

Identification of genes required for segregation of chromosomes in meiosis (scm) is difficult because in most organisms high-fidelity chromosome segregation is essential to produce viable meiotic products. The biology of fission yeast Schizosaccharomyces pombe facilitates identification of such genes. Insertional mutagenesis was achieved by electroporation of linear ura4+ DNA into cells harboring a ura4 deletion. Approximately 1000 stable transformants were screened individually for the production of elevated frequencies of aneuploid spore colonies. Twenty-two candidates were subjected to a secondary screen for cytological defects. Five mutants exhibited significant levels of aberrant meiotic chromosome segregation, but were proficient for mating and completion of meiosis. Each mutant's phenotype cosegregated with its respective ura4+ transgene. The mutations were recessive and defined five complementation groups, revealing five distinct genes (scm1, scm2, scm3, scm4 and scm5). Southern blotting revealed single-site integration in each transformant, indicating that insertional mutagenesis is useful for generating single-locus scm mutations linked to a selectable marker. The transgene insertion points were refractory to analysis by inverse-PCR. Molecular and real-time PCR analyses revealed the presence of multiple, truncated copies of ura4+ at each integration site. Thus, electroporation-mediated insertional mutagenesis in S.pombe is preceded by exonucleolytic processing and concatomerization of the transforming DNA.

The Iml3 protein of the budding yeast is required for the prevention of precocious sister chromatid separation in meiosis I and for sister chromatid disjunction in meiosis II

The mitotic kinetochore of the budding yeast contains a number of proteins which are required for chromosome transmission but are non-essential for vegetative growth. We show that one such protein, Iml3, is essential for meiosis, in that the absence of this protein results in reduced spore viability, precocious sister chromatid segregation of artificial and natural chromosomes in meiosis I and chromosome non-disjunction in meiosis II.