The frequency of X-chromosome nondisjunction in females with lethal
Abstract
Chromosome nondisjunction in meiosis causes the gene disbalance and a number of anomalies in development and fertility. Otherwise, genetically programmed sex-ratio meiotic drive occurs in a number of species. One of the forms of eukaryotic genome organization is a chromocenter evolutionally involved in the regulation of chromosome behavior in dividing cells among insects, plants, mammals, mollusks, and even yeast. In Drosophila, TBP related factor 2 (Trf2) belongs to a conservative Tbp (TATA box-binding protein) gene family and encodes a basic transcription factor. Recent data demonstrates that a decrease in TRF2 expression can result in the abnormalities of chromatin condensation; however, no details of this process have been studied. We demonstrated that a decrease in the TRF2 expression damaged proper chromocenter structure and abolished chromatin condensation and it was a reason for the chromosome nondisjunction. We found that compact chromocenter and correct homologue pairing were abolished in flies with a lower Trf2 expression in germline and in somatic cells. We conclude that TRF2 can not only be involved in transcription activation, but also may perform structural function in pericentromeric heterochromatin organization. The possibility of TRF2 to regulate the evolutionary genetically programmed sex-ratio meiotic drive is discussed.
Keywords
- chromocenter
- chromosome nondisjunction
- asinapsis
- TBP-related factor 2
- Drosophila
1. Introduction
Chromosome nondisjunction during meiosis causes the gene disbalance and, consequently, a number of anomalies in development and fertility. On the other hand, genetically programmed sex-ratio meiotic drive occurs in a number of animal species when mainly males or females are born, which is normal within the given species [1]. The genetic regulation of these processes is actively being studied. There are many factors that can result in the incorrect chromosome segregation. The correct segregation of sister chromatids between daughter cells depends on the coordinated interaction of centrosomes, centromeres, kinetochores, spindle fibrils, topoisomerases, proteolytic processes, and motor proteins [2]. On the other hand, chromosomes must be “prepared” (or structurally organized) when they enter meiosis (or mitosis). Structural disorganization of chromosome or same their regions that control the correct pairing of homologs during meiosis frequently results in the incorrect chromosome segregation. The one way of eukaryotic genome organization is chromocenter, which is evolutionally involved in the regulation of chromosome behavior in dividing cells not only among insects but also among plants, mammals, mollusks, and even yeast [3–7]. This nuclear structure arises in differentiated somatic and germ cells during interphase and meiotic prophase. The chromocenter is generated by the association of pericentromeric regions of all or separate groups of chromosomes and plays an important role in spatial organization of chromosomes [8]. Studies on
In
In the previous studies, we demonstrated that the
Recent data demonstrated that a decrease in
Data of genetic experiments for the analysis of the frequency of X-chromosome nondisjunction in mutant lines and of cytogenetic experiments studding the structure of chromosomes in germ and somatic cells are presented below.
2. Analysis of frequency of X-chromosome nondisjunction in lines with lethal Trf2 mutations
We calculated frequencies of X-chromosome nondisjunction in two groups of lines that contain lethal
The lines of second group were obtained in our laboratory:
In lines with lethal mutation, the X chromosome is maintained on the
When X-chromosome nondisjunction occurred, males and females of exceptional classes (that always differ phenotypically) were detected in descendants. These were X/0 males with normal oval eyes and yellow bodies and XX/Y females with grey bodies and kidney-shaped eyes. Males of the normal class hemizygous for the X chromosome with a lethal allele—
To estimate the influence of the
To determine the influence of
All experiments were repeated three times, and the average frequency of X-chromosome nondisjunction Δ
Alleles | Normal classes | Exceptional classes | |||
---|---|---|---|---|---|
379 | 328 | 13 | 44 | 13.9 | |
296 | 240 | 14 | 15 | 9.8 | |
345 | 328 | 13 | 38 | 13.2 | |
454 | 348 | 28 | 16 | 9.9 | |
306 | 262 | 57 | 72 | 31.2 | |
377 | 342 | 46 | 47 | 20.6 | |
324 | 258 | 14 | 19 | 10.2 | |
345 | 298 | 4 | 15 | 5.6 | |
183 | 218 | 6 | 6 | 5.6 | |
450 | 402 | 10 | 52 | 12.7 | |
478 | 530 | 32 | 6 | 7.0 | |
290 | 268 | 8 | 14 | 7.3 | |
220 | 224 | 26 | 54 | 26.5 | |
361 | 292 | 6 | 8 | 4.1 | |
450 | 306 | 20 | 14 | 8.3 | |
427 | 304 | 34 | 34 | 15.7 | |
439 | 334 | 12 | 8 | 4.9 | |
466 | 360 | 6 | 20 | 5.9 | |
1242 | 1186 | 6 | 11 | 1.4 | |
363 | 282 | 1 | 4 | 1.5 | |
356 | 320 | 1 | 3 | 1.2 |
3. Study of the origin of chromosome nondisjunction in lawc mutants
To identify the source of chromosome nondisjunction, we decided to study the meiosis of mutant females. We performed the cytological analysis of the oocyte nucleus in mutant
In germarium, the oocyte passes through the premeiotic DNA replication, meiosis prophase I, prometaphase I, and metaphase I. In mature oocyte of stage 14, division arrest usually occurs at the stage of metaphase I; chromosomes are collected in karyosome; and only achiasmatic chromosomes (IV and rarely X chromosome) are already oriented to opposite poles (Figure 2A).
We found that in anaphase I the chromocenter in mutant oocyte was often split and the compact karyosome structure was often broken (Figure 2B). The split karyosome assumes the disruption of the chromocenter; therefore, we performed an analysis of the early oocyte at the stage of meiosis prophase I when oocyte chromosomes were held together by pericentromeric heterochromatin, and the compact chromocenter was easy to distinguish. As a result, we found that chromosome compaction and homolog pairing were disturbed in mutant females, and the splitting of the chromocenter was proved to exist (Figure 2C and D).
Thus, a decreasing of
4. Trf2 participates in pericentromeric heterochromatin formation
Chromocenter splitting assumes the disruption of interchromosomal ectopic contacts in the pericentromeric heterochromatin region. We decided to examine
As the
5. The effect of Trf2 knockdown on salivary glands polytene chromosome morphology
We demonstrated in the above described experiments that the decrease in
As considered, polytene chromosomes are very favor objects for the analysis of numerous features of interphase chromosome organization and the genome as a whole [24]. To confirm our hypothesis, we decided to use
For specific
Normally, polytene chromosomes are present in salivary glands in singular due to the somatic synapsis occurs when two homologous chromosomes remain consistently conjugated. Polytene nonhomologous chromosomes in the nucleus are joined by their centromeres to form the most compact common region—chromocenter (Figure 4A, C, and E). Studies of
It is known that partial asynapsis is not a consequence of squashing of nuclei and variations in methods used to make preparations do not affect the frequencies of asynapsis [26]. So, we concluded that high frequency of chromosome asynapsis was induced by
It is known that the chromocenter is responsible for the chromosome co-orientation during cell division and facilitates the paring of homologs [9, 27]. The disturbance of paring affects the transvection (or allelic complementation)—the phenomenon in which gene regulatory elements located in one of the homologs control a promoter of the same gene but located in another homolog [28, 29]. It is interesting to note that hypomorphic
The study a set of mutations that cause chromosome nondisjunction allowed to conclude that the chromocenter is a genetically programmed structure, that is, there are genes that control its formation and reorganization [11]. For example, it was demonstrated that the recessive mutation of
The
Another gene—
It was shown that
In yeast, it was demonstrated that kinetochores—large protein complexes assembled on the centromeric region of the chromosomes, to which spindle microtubule is attached during cell division—are formed by the epigenetic mechanism. This mechanism involves the generation of specialized nucleosomes in which a canonical histone H3 is replaced by its centromere-specific homologs—centromere protein A (CENP-A). This protein served as a landmark for kinetochore assembly to define the identity of centromeres [37, 38]. The high frequency of chromosome nondisjunction induced by decondensation of pericentric heterochromatin in
As it was mentioned above, the correct distribution of chromatids between daughter cells depends on the coordinated interaction of centrosomes, centromeres, kinetochores, spindle fibrils, topoisomerase, proteolytic processes, and motor proteins. The error of accurate spatiotemporal interactions between any of these factors results in a genomic disbalance. We cannot completely exclude the probability that
Process | Genes | |
---|---|---|
Chromatin compaction | ||
Assembly of division spindle | ||
Chromosome disjunction | ||
Organization of actin components of cytoskeleton | ||
Checkpoint | ||
? |
This does not mean that
6. Conclusion
We demonstrated that a decrease in the
In conclusion, we would like to note that in the recent screening for genes that control the sex-ratio meiotic drive in
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