RESUMO
X chromosome dosage compensation ensures balanced gene dosage between the X chromosome and autosomes and between the sexes, involving divergent mechanisms among mammals. We elucidated a distinct mechanism for X chromosome inactivation (XCI) in cynomolgus monkeys, a model for human development. The trophectoderm and cytotrophoblast acquire XCI around implantation through an active intermediate bearing repressive modifications and compacted structure, whereas the amnion, epiblast, and hypoblast maintain such an intermediate protractedly, attaining XCI by a week after implantation. Males achieve X chromosome up-regulation (XCU) progressively, whereas females show XCU coincidentally with XCI, both establishing the X:autosome dosage compensation by 1 week after implantation. Conversely, primordial germ cells undergo X chromosome reactivation by reversing the XCI pathway early during their development. Our findings establish a foundation for clarifying the dosage compensation mechanisms in primates, including humans.
Assuntos
Blastocisto/fisiologia , Mecanismo Genético de Compensação de Dose , Macaca fascicularis/embriologia , Macaca fascicularis/genética , Trofoblastos/fisiologia , Inativação do Cromossomo X , Cromossomo X/genética , Animais , Desenvolvimento Embrionário , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genes Ligados ao Cromossomo X , Células Germinativas/fisiologia , Histonas/metabolismo , Metilação , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Regulação para Cima , Cromossomo X/metabolismo , Cromossomo X/ultraestruturaRESUMO
Diverse cell types in tissues have distinct gene expression programs, chromatin states, and nuclear architectures. To correlate such multimodal information across thousands of single cells in mouse brain tissue sections, we use integrated spatial genomics, imaging thousands of genomic loci along with RNAs and epigenetic markers simultaneously in individual cells. We reveal that cell typespecific association and scaffolding of DNA loci around nuclear bodies organize the nuclear architecture and correlate with differential expression levels in different cell types. At the submegabase level, active and inactive X chromosomes access similar domain structures in single cells despite distinct epigenetic and expression states. This work represents a major step forward in linking single-cell three-dimensional nuclear architecture, gene expression, and epigenetic modifications in a native tissue context.
Assuntos
Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , Córtex Cerebral/citologia , Neuroglia/ultraestrutura , Neurônios/ultraestrutura , Análise de Célula Única , Animais , Córtex Cerebral/metabolismo , Cromatina/metabolismo , Cromatina/ultraestrutura , Cromossomos/metabolismo , Cromossomos/ultraestrutura , Células Endoteliais/metabolismo , Células Endoteliais/ultraestrutura , Epigênese Genética , Feminino , Genoma , Hibridização in Situ Fluorescente , Camundongos , Neuroglia/metabolismo , Neurônios/metabolismo , RNA-Seq , Transcrição Gênica , Transcriptoma , Cromossomo X/metabolismo , Cromossomo X/ultraestruturaRESUMO
Chromosome structural change has long been considered important in the evolution of post-zygotic reproductive isolation. The premise that karyotypic variation can serve as a possible barrier to gene flow is founded on the expectation that heterozygotes for structurally distinct chromosomal forms would be partially sterile (negatively heterotic) or show reduced recombination. We report the outcome of a detailed comparative molecular cytogenetic study of three antelope species, genus Raphicerus, that have undergone a rapid radiation. The species are largely conserved with respect to their euchromatic regions but the X chromosomes, in marked contrast, show distinct patterns of heterochromatic amplification and localization of repeats that have occurred independently in each lineage. We argue a novel hypothesis that postulates that the expansion of heterochromatic blocks in the homogametic sex can, with certain conditions, contribute to post-zygotic isolation. i.e., female hybrid incompatibility, the converse of Haldane's rule. This is based on the expectation that hybrids incur a selective disadvantage due to impaired meiosis resulting from the meiotic checkpoint network's surveillance of the asymmetric expansions of heterochromatic blocks in the homogametic sex. Asynapsis of these heterochromatic regions would result in meiotic silencing of unsynapsed chromatin and, if this persists, germline apoptosis and female infertility.
Assuntos
Antílopes/genética , Especiação Genética , Cariótipo , Modelos Genéticos , Isolamento Reprodutivo , Cromossomo X/ultraestrutura , África , Animais , Antílopes/classificação , Feminino , Fluxo Gênico , Heterozigoto , Hibridização Genética , Hibridização in Situ Fluorescente , Infertilidade Feminina/genética , Masculino , Meiose , Recombinação Genética , Fatores SexuaisRESUMO
The genus Psammolestes within the subfamily Triatominae and tribe Rhodniini comprises the species Psammolestes arthuri, Psammolestes coreodes, and Psammolestes tertius, all potential vectors of Chagas disease. A feature of Psammolestes is their close association with birds, which makes them an interesting model for evolutionary studies. We analyzed cytogenetically Psammolestes spp., with the aim of contributing to the genetic and evolutionary knowledge of these vectors. All species of the Psammolestes showed the same chromosomal characteristics: chromocenter formed only by sex chromosomes X and Y, karyotype 2n = 22 and constitutive heterochromatin, and AT base pairs restricted to the sex chromosome Y. These results corroborate the monophyly of the genus and lead to the hypothesis that during the derivation of P. tertius, P. coreodes, and P. arthuri from their common ancestor, there was no reorganization in the number or structure of chromosomes.
Assuntos
Doença de Chagas/transmissão , Cromossomos de Insetos/química , Especiação Genética , Insetos Vetores/genética , Filogenia , Triatominae/genética , Animais , Pareamento de Bases , Aves/parasitologia , Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , Cromossomos de Insetos/ultraestrutura , Heterocromatina/química , Heterocromatina/ultraestrutura , Humanos , Insetos Vetores/classificação , Insetos Vetores/parasitologia , Cariótipo , América Latina/epidemiologia , Triatominae/classificação , Triatominae/parasitologia , Trypanosoma cruzi/patogenicidade , Cromossomo X/química , Cromossomo X/ultraestrutura , Cromossomo Y/química , Cromossomo Y/ultraestruturaRESUMO
Heteromorphic sex chromosomes are common in eukaryotes and largely ubiquitous in birds and mammals. The largest number of multiple sex chromosomes in vertebrates known today is found in the monotreme platypus (Ornithorhynchus anatinus, 2n = 52) which exhibits precisely 10 sex chromosomes. Interestingly, fish, amphibians, and reptiles have sex determination mechanisms that do or do not involve morphologically differentiated sex chromosomes. Relatively few amphibian species carry heteromorphic sex chromosomes, and when present, they are frequently represented by only one pair, either XX:XY or ZZ:ZW types. Here, in contrast, with several evidences, from classical and molecular cytogenetic analyses, we found 12 sex chromosomes in a Brazilian population of the smoky jungle frog, designated as Leptodactylus pentadactylus Laurenti, 1768 (Leptodactylinae), which has a karyotype with 2n = 22 chromosomes. Males exhibited an astonishing stable ring-shaped meiotic chain composed of six X and six Y chromosomes. The number of sex chromosomes is larger than the number of autosomes found, and these data represent the largest number of multiple sex chromosomes ever found among vertebrate species. Additionally, sequence and karyotype variation data suggest that this species may represent a complex of species, in which the chromosomal rearrangements may possibly have played an important role in the evolution process.
Assuntos
Anuros/genética , Processos de Determinação Sexual , Cromossomo X/metabolismo , Cromossomo Y/metabolismo , Animais , Anuros/classificação , Brasil , Hibridização Genômica Comparativa , Feminino , Hibridização in Situ Fluorescente , Cariótipo , Cariotipagem , Masculino , Filogenia , Cromossomo X/ultraestrutura , Cromossomo Y/ultraestruturaRESUMO
X inactivation is a fundamental mechanism in eutherian mammals to restore a balance of X-linked gene products between XY males and XX females. However, it has never been extensively studied in a eutherian species with a sex determination system that deviates from the ubiquitous XX/XY. In this study, we explore the X inactivation process in the African pygmy mouse Mus minutoides, that harbours a polygenic sex determination with three sex chromosomes: Y, X, and a feminizing mutant X, named X*; females can thus be XX, XX*, or X*Y, and all males are XY. Using immunofluorescence, we investigated histone modification patterns between the two X chromosome types. We found that the X and X* chromosomes are randomly inactivated in XX* females, while no histone modifications were detected in X*Y females. Furthermore, in M. minutoides, X and X* chromosomes are fused to different autosomes, and we were able to show that the X inactivation never spreads into the autosomal segments. Evaluation of X inactivation by immunofluorescence is an excellent quantitative procedure, but it is only applicable when there is a structural difference between the two chromosomes that allows them to be distinguished.
Assuntos
Cromossomos de Mamíferos/metabolismo , Processamento de Proteína Pós-Traducional , Processos de Determinação Sexual , Inativação do Cromossomo X , Cromossomo X/metabolismo , Cromossomo Y/metabolismo , Acetilação , África , Animais , Animais Selvagens , Cromossomos de Mamíferos/ultraestrutura , Feminino , Imunofluorescência/métodos , Histonas/genética , Histonas/metabolismo , Cariotipagem , Masculino , Metilação , Camundongos , Cromossomo X/ultraestrutura , Cromossomo Y/ultraestruturaRESUMO
A striking difference between male and female nuclei was recognized early on by the presence of a condensed chromatin body only in female cells. Mary Lyon proposed that X inactivation or silencing of one X chromosome at random in females caused this structural difference. Subsequent studies have shown that the inactive X chromosome (Xi) does indeed have a very distinctive structure compared to its active counterpart and all autosomes in female mammals. In this review, we will recap the discovery of this fascinating biological phenomenon and seminal studies in the field. We will summarize imaging studies using traditional microscopy and super-resolution technology, which revealed uneven compaction of the Xi. We will then discuss recent findings based on high-throughput sequencing techniques, which uncovered the distinct three-dimensional bipartite configuration of the Xi and the role of specific long non-coding RNAs in eliciting and maintaining this structure. The relative position of specific genomic elements, including genes that escape X inactivation, repeat elements and chromatin features, will be reviewed. Finally, we will discuss the position of the Xi, either near the nuclear periphery or the nucleolus, and the elements implicated in this positioning.This article is part of the themed issue 'X-chromosome inactivation: a tribute to Mary Lyon'.
Assuntos
Cromatina/metabolismo , RNA Longo não Codificante/metabolismo , Inativação do Cromossomo X/genética , Cromossomo X/genética , Animais , Cromossomos Humanos X/genética , Cromossomos Humanos X/ultraestrutura , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Camundongos , Cromossomo X/ultraestruturaRESUMO
The heteromorphic X and Y chromosomes behave in a special way in mammalian spermatocytes; they form the XY body and synapse only partially. The aim of this article was to study the origin and the role of the special differentiations in the XY pair of the domestic cat during pachytene by analyzing its fine structural characteristics and the immunolocalization of the main meiotic proteins SYCP3, SYCP1, SYCE3, SMC3, γ-H2AX, BRCA1, H3K27me3, and MLH1. The cat XY body shows particularly striking structures: an extreme degree of axial fibrillation in late pachynema and a special location of SYCP3-containing fibrils, bridging different regions of the main X axis, as well as one bridge at the inner end of the pairing region that colocalizes with the single mandatory MLH1 focus. There are sequential changes, first bullous expansions, then subdivision into fibrils, all involving axial thickening. The chromatin of the XY body presents the usual features of meiotic sex chromosome inactivation. An analysis of the XY body of many eutherians and metatherians suggests that axial thickenings are primitive features. The sequential changes in the mass and location of SYCP3-containing fibers vary among the clades because of specific processes of axial assembly/disassembly occurring in different species.
Assuntos
Gatos/genética , Proteínas Nucleares/metabolismo , Estágio Paquíteno/genética , Complexo Sinaptonêmico/metabolismo , Cromossomo X/metabolismo , Cromossomo X/ultraestrutura , Cromossomo Y/metabolismo , Cromossomo Y/ultraestrutura , Animais , Proteína BRCA1/genética , Proteína BRCA1/metabolismo , Cromatina/metabolismo , Cromatina/ultraestrutura , Histonas/genética , Histonas/metabolismo , Masculino , Microscopia de Fluorescência , Proteína 1 Homóloga a MutL/genética , Proteína 1 Homóloga a MutL/metabolismo , Espermatócitos/metabolismo , Complexo Sinaptonêmico/genéticaRESUMO
We show that the geophilomorph centipede Strigamia maritima possesses an XX/XY system of sex chromosomes, with males being the heterogametic sex. This is, to our knowledge, the first report of sex chromosomes in any geophilomorph centipede. Using the recently assembled Strigamia genome sequence, we identified a set of scaffolds differentially represented in male and female DNA sequence. Using quantitative real-time PCR, we confirmed that three candidate X chromosome-derived scaffolds are present at approximately twice the copy number in females as in males. Furthermore, we confirmed that six candidate Y chromosome-derived scaffolds contain male-specific sequences. Finally, using this molecular information, we designed an X chromosome-specific DNA probe and performed fluorescent in situ hybridization against mitotic and meiotic chromosome spreads to identify the Strigamia XY sex-chromosome pair cytologically. We found that the X and Y chromosomes are recognizably different in size during the early pachytene stage of meiosis, and exhibit incomplete and delayed pairing.
Assuntos
Artrópodes/genética , Processos de Determinação Sexual/genética , Cromossomo X , Cromossomo Y , Animais , Artrópodes/embriologia , Artrópodes/crescimento & desenvolvimento , Artrópodes/fisiologia , Hibridização Genômica Comparativa , Feminino , Hibridização in Situ Fluorescente , Masculino , Reação em Cadeia da Polimerase em Tempo Real , Cromossomo X/ultraestrutura , Cromossomo Y/ultraestruturaRESUMO
Trisomy, the presence of a third copy of one chromosome, is deleterious and results in inviable or defective progeny if passed through the germ line. Random segregation of an extra chromosome is predicted to result in a high frequency of trisomic offspring from a trisomic parent. Caenorhabditis elegans with trisomy of the X chromosome, however, have far fewer trisomic offspring than expected. We found that the extra X chromosome was preferentially eliminated during anaphase I of female meiosis. We utilized a mutant with a specific defect in pairing of the X chromosome as a model to investigate the apparent bias against univalent inheritance. First, univalents lagged during anaphase I and their movement was biased toward the cortex and future polar body. Second, late-lagging univalents were frequently captured by the ingressing polar body contractile ring. The asymmetry of female meiosis can thus partially correct pre-existing trisomy.
Assuntos
Divisão Celular Assimétrica/genética , Caenorhabditis elegans/genética , Padrões de Herança , Trissomia , Cromossomo X/química , Anáfase , Animais , Segregação de Cromossomos , Feminino , Oócitos/metabolismo , Oócitos/ultraestrutura , Cromossomo X/ultraestruturaRESUMO
In mammals, the process of X-chromosome inactivation ensures equivalent levels of X-linked gene expression between males and females through the silencing of one of the two X chromosomes in female cells. The process is established early in development and is initiated by a unique locus, which produces a long noncoding RNA, Xist. The Xist transcript triggers gene silencing in cis by coating the future inactive X chromosome. It also induces a cascade of chromatin changes, including posttranslational histone modifications and DNA methylation, and leads to the stable repression of all X-linked genes throughout development and adult life. We review here recent progress in our understanding of the molecular mechanisms involved in the initiation of Xist expression, the propagation of the Xist RNA along the chromosome, and the cis-elements and trans-acting factors involved in the maintenance of the repressed state. We also describe the diverse strategies used by nonplacental mammals for X-chromosome dosage compensation and highlight the common features and differences between eutherians and metatherians, in particular regarding the involvement of long noncoding RNAs.
Assuntos
Inativação Gênica , RNA Longo não Codificante/genética , Inativação do Cromossomo X/genética , Animais , Cromatina/genética , Cromatina/ultraestrutura , Mapeamento Cromossômico , Cromossomos Humanos X/genética , Células-Tronco Embrionárias/ultraestrutura , Evolução Molecular , Feminino , Impressão Genômica , Humanos , Elementos Nucleotídeos Longos e Dispersos , Masculino , Marsupiais/genética , Camundongos , Processos de Determinação Sexual , Fatores de Transcrição/genética , Cromossomo X/genética , Cromossomo X/ultraestruturaRESUMO
The XY body from spermatocytes of the rodent Galea musteloides shows progressive changes of the synaptonemal complex (SC) axes and the X-chromatin during pachynema. There is a gross thickening of the X-axis and the formation of a large X chromosome loop at mid and late pachytene stages. The SC proteins synaptonemal complex protein 3 (SYCP3), synaptonemal complex protein 1, and synaptonemal complex central element protein 3 and the proteins breast cancer 1, MutL homolog 1 (MLH1), and radiation-repair 51 (related to meiotic processes), the cohesin structural maintenance of chromosome 3, the centromeric protein (with CREST antibody), and the silenced chromatin (with phosphorylated (139ph) H2A histone family, member X (γ-H2AX) antibody) were analyzed in this XY body. The thick X-axis, including the interstitial loop, becomes formed by four to six laminae showing a cross-striation with a periodicity of about 20 nm. The whole length of the gross X-axis shows no significant changes during pachynema, but the interstitial chromatin of the X chromosome and the X centromere are included in the large loop, and it becomes separated from the SC. A conventional SC formed by the Y-axis, a central region and a thin lateral element originally corresponding to the X-axis, remains undisturbed up to the end of pachynema. A single MLH1 focus develops either at the distal or the proximal region of the loop end attached to the conventional SC. The chromatin surrounding the thickened axis is labeled with γ-H2AX. It is shown that most of the SYCP3 protein associated with the X chromosome loop is not involved in the SC maintenance, but it is located with the cohesin axis separated from the SC proper.
Assuntos
Roedores/genética , Complexo Sinaptonêmico/ultraestrutura , Cromossomo X/ultraestrutura , Animais , Centrômero/genética , Centrômero/ultraestrutura , Cromatina/genética , Cromatina/ultraestrutura , Evolução Molecular , Inativação Gênica , Cobaias , Masculino , Membrana Nuclear , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Estágio Paquíteno , Análise de Sequência de DNA , Complexo Sinaptonêmico/genética , Cromossomo X/genéticaRESUMO
Many large noncoding RNAs (lncRNAs) regulate chromatin, but the mechanisms by which they localize to genomic targets remain unexplored. We investigated the localization mechanisms of the Xist lncRNA during X-chromosome inactivation (XCI), a paradigm of lncRNA-mediated chromatin regulation. During the maintenance of XCI, Xist binds broadly across the X chromosome. During initiation of XCI, Xist initially transfers to distal regions across the X chromosome that are not defined by specific sequences. Instead, Xist identifies these regions by exploiting the three-dimensional conformation of the X chromosome. Xist requires its silencing domain to spread across actively transcribed regions and thereby access the entire chromosome. These findings suggest a model in which Xist coats the X chromosome by searching in three dimensions, modifying chromosome structure, and spreading to newly accessible locations.
Assuntos
Genoma , RNA Longo não Codificante/metabolismo , Inativação do Cromossomo X , Cromossomo X/metabolismo , Animais , Diferenciação Celular , Linhagem Celular , Cromatina/química , Cromatina/metabolismo , Feminino , Masculino , Camundongos , Modelos Genéticos , RNA Longo não Codificante/química , Transcrição Gênica , Cromossomo X/ultraestruturaRESUMO
Three xenarthrans species Chaetophractus villosus, Chaetophractus vellerosus, and Zaedyus pichiy have been used for the analysis of the structure, behavior, and immunochemical features of the XY body during pachytene. In all these species, the sex chromosomes form an XY body easily identifiable in thin sections by the special and regular packing of the chromatin fibers of the internal region of the XY body ("differential" regions) and those of the peripheral region (synaptic region). Spermatocyte spreads show a complete synapsis between the X- and the Y-axis, which lasts up to the end of pachytene. From the early pachytene substages to the late ones, the X-axis develops prominent branches, which in late pachytene span the synaptic region. Synapsis is regular as shown by SYCP1 labeling. Axial development is followed by SYCP3 labeling and in the asynaptic region of the X-axis by BRCA1. Gamma-H2AX labels exclusively the differential (asynaptic) region of the X chromosome. A single focus is labeled by MLH1 in the synaptic region. The location of this MLH1 focus spans from 0.3 to 1.6 µm from the telomere in the analyzed xenarthrans, covering approximately half of the Y-axis length. It is concluded that xenarthrans, as basal placental mammals, harbor the largest pseudoautosomal regions of presently analyzed mammals, and shows the typical features of meiotic sex chromosome inactivation (MSCI).
Assuntos
Tatus/genética , Montagem e Desmontagem da Cromatina , Pareamento Cromossômico , Recombinação Genética , Animais , Cromatina/ultraestrutura , Masculino , Cromossomos Sexuais , Espermatócitos/metabolismo , Cromossomo X/ultraestrutura , Cromossomo Y/ultraestruturaRESUMO
In many eutherian mammals, X-Y chromosome pairing and recombination is required for meiotic progression and correct sex chromosome disjunction. Arvicoline rodents present a notable exception to this meiotic rule, with multiple species possessing asynaptic sex chromosomes. Most asynaptic vole species belong to the genus Microtus sensu lato. However, many of the species both inside and outside the genus Microtus display normal X-Y synapsis at meiosis. These observations suggest that the synaptic condition was present in the common ancestor of all voles, but gaps in current taxonomic sampling across the arvicoline phylogeny prevent identification of the lineage(s) along which the asynaptic state arose. In this study, we use electron and immunofluorescent microscopy to assess heterogametic sex chromosome pairing in 12 additional arvicoline species. Our sample includes ten species of the tribe Microtini and two species of the tribe Lagurini. This increased breadth of sampling allowed us to identify asynaptic species in each major Microtine lineage. Evidently, the ability of the sex chromosomes to pair and recombine in male meiosis has been independently lost at least three times during the evolution of Microtine rodents. These results suggest a lack of evolutionary constraint on X-Y synapsis in Microtini, hinting at the presence of alternative molecular mechanisms for sex chromosome segregation in this large mammalian tribe.
Assuntos
Arvicolinae/genética , Pareamento Cromossômico , Meiose/genética , Cromossomo X , Cromossomo Y , Animais , Masculino , Espermatócitos/metabolismo , Cromossomo X/ultraestrutura , Cromossomo Y/ultraestruturaRESUMO
Fissioncytorace-1, a member of the nasuta-albomicans complex of Drosophila is an evolutionary product of centric fission, which had occurred in the chromosome X3 of Cytorace 1, a hydridization product of Drosophila nasuta nasuta male (2n=8) and Drosophila nasuta albomicans female (2n=6). Cytorace 1 (males 2n=7; females 2n=6) has inherited this chromosome from its D. n. albomicans parent. The chromosome X3 of D. n. albomicans is a derivative of a centric fusion between the acrocentric chromosome 3 and the chromosome X of D. n. nasuta. The Fissioncytorace-1 has crossed 200 generations from the time of its evolution in the laboratory environment. When this centromeric fission race was subjected to some of the morphophenotypic and fitness assessment to find its overall population fitness showed, increased body size, sternopleural bristle, ovarioles, lifetime fecundity and fertility with reduced interspecific competitive ability and hatching success when compared with its parent (Cytorace 1). These results suggest that the hybrid races must have encountered an early event of recombinational raciation during their evolution in the laboratory environment, which is a unique observation in animal system illustrating the increase in the tempo of evolution after the event of hybridization.
Assuntos
Centrômero/ultraestrutura , Evolução Molecular Direcionada , Drosophila/classificação , Animais , Tamanho Corporal/genética , Cromossomos/ultraestrutura , Drosophila/genética , Drosophila/fisiologia , Feminino , Fertilidade/genética , Aptidão Genética/genética , Cariotipagem , Masculino , Ovário/ultraestrutura , Cromossomo X/ultraestruturaRESUMO
Karyotypes can be difficult to create due to odd chromosome behaviors and high levels of adhesion between sex chromosomes. We have used live-cell imaging and micromanipulation to determine precisely the karyotype of the orb-weaving spider Neoscona arabesca (Walckenaer). We have found that N. arabesca has a sex determination mechanism of XO (male)-XX (female) with 2n = 20 autosomes. Staining of female tissues revealed a chromosome number of 22 in mitotic cells.
Assuntos
Processos de Determinação Sexual , Aranhas/genética , Cromossomo X/genética , Animais , Bandeamento Cromossômico , Feminino , Cariotipagem , Masculino , Micromanipulação , Mitose , Cromossomo X/ultraestruturaRESUMO
The effect of ectopic expression of male specific lethal 2 (msl2) on chromatin modification and gene expression was studied in Drosophila diploid females and metafemales (3X;2A). Results show that ectopic expression of MSL2 in transgenic msl2 females and metafemales sequesters the MOF histone acetylase to the X, which occurs concordantly with an increase of histone acetylation. Gene expression studies indicate that the X-linked genes are not affected by direct targeting of the MSL complex and the resulting increased H4Lys16 acetylation on the X chromosomes, suggesting one function of the MSL complex is to nullify the effect of a high level of histone acetylation. These results are not consistent with the hypothesis that the presence of the MSL complex conditions a two-fold upregulation. Autosomal gene expression is generally decreased in ectopically expressed MSL2 females, which correlates with the reduced autosomal histone acetylation. Metafemales show dosage compensation of X-linked genes with some autosomal reductions in expression. Interestingly, in metafemales with ectopically expressed MSL2, the autosomal expression is returned to a more normal level. There is a lower autosomal level of histone acetylation compared to the normal metafemales, suggesting a nullifying effect on the negative dosage effect of the X chromosome as previously hypothesized to occur in normal males.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Mecanismo Genético de Compensação de Dose , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Histona Acetiltransferases/metabolismo , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/ultraestrutura , Feminino , Expressão Gênica/fisiologia , Genes Ligados ao Cromossomo X/fisiologia , Histona Acetiltransferases/genética , Masculino , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Cromossomo X/metabolismo , Cromossomo X/ultraestruturaRESUMO
Acquisition of knowledge of the nature and DNA content of B chromosomes has been triggered by a collection of molecular techniques, one of which, microdissection, has provided interesting results in a number of B chromosome systems. Here we provide the first data on the molecular composition of B chromosomes in Locusta migratoria, after microdissection of the B and X chromosomes, DNA amplification by one (B) or two (X) different methods, and chromosome painting. The results showed that B chromosomes share at least two types of repetitive DNA sequences with the A chromosomes, suggesting that Bs in this species most likely arose intraspecifically. One of these repetitive DNAs is located on the heterochromatic distal half of the B chromosome and in the pericentromeric regions of about half of the A chromosomes, including the X. The other type of repetitive DNA is located interspersedly over the non-centromeric euchromatic regions of all A chromosomes and in an interstitial part of the proximal euchromatic half of the B chromosome. Chromosome painting, however, did not provide results sufficiently reliable to determine, in this species, which A chromosome gave rise to the B; this might be done by detailed analysis of the microdissected DNA sequences.
Assuntos
Cromossomos/genética , Locusta migratoria/genética , Cromossomo X/genética , Animais , Coloração Cromossômica , Cromossomos/ultraestrutura , DNA/genética , Microdissecção , Estágio Paquíteno , Cromossomo X/ultraestruturaRESUMO
The 3D-FISH was employed to investigate the telomere topology in polytene nuclei of salivary glands of Drosophila melanogaster. The majorities of telomeres in polytene nuclei of salivary glands in Drosophila strain y(2-717) are localized in the nuclear central area and have no contacts with nuclear membrane. In females of this strain, ectopic contacts between telomeres occur at 25 % higher frequency than in males. HeT-A DNA in y(2-717alk3-2) strain, which is a derivative of y(2-717) carrying an inversion between 1D and 13C bands, is found in region 13 of X chromosome. The frequency of ectopic contacts of telomeres in y(2-717alk3-2) males is 10 % higher than that in y(2-717) strain. The number of ectopic contacts can be significantly different in independent experiments, possibly indicating the role of random factors in the contact formation.