RESUMO
The tea plant (Camellia sinensis) is a thermophilic cash crop and contains a highly duplicated and repeat-rich genome. It is still unclear how DNA methylation regulates the evolution of duplicated genes and chilling stress in tea plants. We therefore generated a single-base-resolution DNA methylation map of tea plants under chilling stress. We found that, compared with other plants, the tea plant genome is highly methylated in all three sequence contexts, including CG, CHG and CHH (where H = A, T, or C), which is further proven to be correlated with its repeat content and genome size. We show that DNA methylation in the gene body negatively regulates the gene expression of tea plants, whereas non-CG methylation in the flanking region enables a positive regulation of gene expression. We demonstrate that transposable element-mediated methylation dynamics significantly drives the expression divergence of duplicated genes in tea plants. The DNA methylation and expression divergence of duplicated genes in the tea plant increases with evolutionary age and selective pressure. Moreover, we detect thousands of differentially methylated genes, some of which are functionally associated with chilling stress. We also experimentally reveal that DNA methyltransferase genes of tea plants are significantly downregulated, whereas demethylase genes are upregulated at the initial stage of chilling stress, which is in line with the significant loss of DNA methylation of three well-known cold-responsive genes at their promoter and gene body regions. Overall, our findings underscore the importance of DNA methylation regulation and offer new insights into duplicated gene evolution and chilling tolerance in tea plants.
Assuntos
Camellia sinensis/genética , Metilação de DNA , Elementos de DNA Transponíveis/genética , Evolução Molecular , Genes Duplicados/genética , Genoma de Planta/genética , Camellia sinensis/fisiologia , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , Tamanho do Genoma , Estresse FisiológicoRESUMO
Gene duplication is a primary means to generate genomic novelties, playing an essential role in speciation and adaptation. Particularly in plants, a high abundance of duplicate genes has been maintained for significantly long periods of evolutionary time. To address the manner in which young duplicate genes were derived primarily from small-scale gene duplication and preserved in plant genomes and to determine the underlying driving mechanisms, we generated transcriptomes to produce the expression profiles of five tissues in Arabidopsis thaliana and the closely related species Arabidopsis lyrata and Capsella rubella Based on the quantitative analysis metrics, we investigated the evolutionary processes of young duplicate genes in Arabidopsis. We determined that conservation, neofunctionalization, and specialization are three main evolutionary processes for Arabidopsis young duplicate genes. We explicitly demonstrated the dynamic functionalization of duplicate genes along the evolutionary time scale. Upon origination, duplicates tend to maintain their ancestral functions; but as they survive longer, they might be likely to develop distinct and novel functions. The temporal evolutionary processes and functionalization of plant duplicate genes are associated with their ancestral functions, dynamic DNA methylation levels, and histone modification abundances. Furthermore, duplicate genes tend to be initially expressed in pollen and then to gain more interaction partners over time. Altogether, our study provides novel insights into the dynamic retention processes of young duplicate genes in plant genomes.
Assuntos
Arabidopsis/genética , Evolução Molecular , Duplicação Gênica , Regulação da Expressão Gênica de Plantas , Genes Duplicados/genética , Genoma de Planta/genética , Brassica rapa/genética , Capsella/genética , Perfilação da Expressão Gênica/métodos , Pólen/genética , Especificidade da Espécie , Fatores de TempoRESUMO
Duplicated genes can contribute to the evolution of new functions and they are common in eukaryotic genomes. After duplication, genes can show divergence in their sequence and/or expression patterns. Qualitative complementary expression, or reciprocal expression, is when only one copy is expressed in some organ or tissue types and only the other copy is expressed in others, indicative of regulatory subfunctionalization or neofunctionalization. From analyses of two microarray data sets with 83 different organ types, developmental stages, and cell types in Arabidopsis thaliana, we determined that 30% of whole-genome duplicate pairs and 38% of tandem duplicate pairs show reciprocal expression patterns. We reconstructed the ancestral state of expression patterns to infer that considerably more cases of reciprocal expression resulted from gain of a new expression pattern (regulatory neofunctionalization) than from partitioning of ancestral expression patterns (regulatory subfunctionalization). Pollen was an especially common organ type for expression gain, resulting in contrasting expression of some duplicates in pollen. Many of the gene pairs with reciprocal expression showed asymmetric sequence rate evolution, consistent with neofunctionalization, and the more rapidly evolving copy often showed a more restricted expression pattern. A gene with reciprocal expression in pollen, involved in brassinosteroid signal transduction, has evolved more rapidly than its paralog, and it shows evidence for a new function in pollen. This study indicates the evolutionary importance of reciprocal expression patterns between gene duplicates, showing that they are common, often associated with regulatory neofunctionalization, and may be a factor allowing for retention and divergence of duplicated genes.
Assuntos
Arabidopsis/citologia , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Genes Duplicados/genética , Genes de Plantas/genética , Especificidade de Órgãos/genética , Arabidopsis/anatomia & histologia , Arabidopsis/enzimologia , Sequência de Bases , Simulação por Computador , Evolução Molecular , Variação Genética , Dados de Sequência Molecular , Fosfoproteínas Fosfatases/genética , Pólen/genéticaRESUMO
Catalase is one of the key antioxidant enzymes and it appears to be involved in protection against immune infection and oxidative stress. Here, two catalase cDNAs (ChCat-1 and ChCat-2) were isolated from hemocytes of Crassostrea hongkongensis using SSH and RACE. The full-length cDNAs of ChCat-1 and ChCat-2 are 1913 and 2466 bp in length, encoding proteins of 515 and 511 amino acids, respectively. Multiple alignments of amino acid sequences revealed that both ChCat-1 and ChCat-2 possess several characteristic features of the catalase family of enzymes, including one proximal active site signature, one heme-ligand signature, and three catalytic amino acid residues (His(72), Asn(145) and Tyr(355)). Phylogenetic analysis indicates that these two catalases may share a common ancestral gene and result from a gene duplication event following the divergence of bivalves and gastropods. Constitutive expression of ChCat-1 and ChCat-2 was observed in all tissues studied, with highest levels of expression in gill and muscle, respectively. The expression of both genes was inducible by bacterial infection, and reached the maximum at 8 h (9.0-fold) and 12 h (2.3-fold) post-infection, respectively. Furthermore, both the purified ChCat-1 and ChCat-2 protein displayed a strong catalase activity, and S2 cells carrying ChCat-1 or ChCat-2 showed a higher degree of resistance to H(2)O(2) than that of control cells. In a word, this is the first report of the presence of two catalase genes in a single marine bivalve, and our results highlight the involvement of both ChCat-1 and ChCat-2 in host protection against pathogen infection and oxidative stress in C. hongkongensis.
Assuntos
Catalase/genética , Catalase/imunologia , Crassostrea/imunologia , Regulação da Expressão Gênica/imunologia , Estresse Oxidativo/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Catalase/metabolismo , Crassostrea/enzimologia , Crassostrea/microbiologia , Primers do DNA/genética , DNA Complementar/genética , Genes Duplicados/genética , Brânquias/metabolismo , Hemócitos/metabolismo , Peróxido de Hidrogênio , Dados de Sequência Molecular , Músculos/metabolismo , Filogenia , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
WRINKLED1 (WRI1), a key regulator of seed oil biosynthesis in Arabidopsis (Arabidopsis thaliana), was duplicated during the genome amplification of the cereal ancestor genome 90 million years ago. Both maize (Zea mays) coorthologs ZmWri1a and ZmWri1b show a strong transcriptional induction during the early filling stage of the embryo and complement the reduced fatty acid content of Arabidopsis wri1-4 seeds, suggesting conservation of molecular function. Overexpression of ZmWri1a not only increases the fatty acid content of the mature maize grain but also the content of certain amino acids, of several compounds involved in amino acid biosynthesis, and of two intermediates of the tricarboxylic acid cycle. Transcriptomic experiments identified 18 putative target genes of this transcription factor, 12 of which contain in their upstream regions an AW box, the cis-element bound by AtWRI1. In addition to functions related to late glycolysis and fatty acid biosynthesis in plastids, the target genes also have functions related to coenzyme A biosynthesis in mitochondria and the production of glycerol backbones for triacylglycerol biosynthesis in the cytoplasm. Interestingly, the higher seed oil content in ZmWri1a overexpression lines is not accompanied by a reduction in starch, thus opening possibilities for the use of the transgenic maize lines in breeding programs.
Assuntos
Regulação da Expressão Gênica de Plantas , Genes Duplicados/genética , Genes de Plantas/genética , Óleos de Plantas/metabolismo , Proteínas de Plantas/genética , Sementes/genética , Zea mays/genética , Arabidopsis/genética , Sequência de Bases , Ácidos Graxos/metabolismo , Perfilação da Expressão Gênica , Teste de Complementação Genética , Glicólise/genética , Modelos Biológicos , Dados de Sequência Molecular , Mutação/genética , Filogenia , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Triglicerídeos/biossínteseRESUMO
Genetic incompatibility is a barrier contributing to species isolation and is caused by genetic interactions. We made a whole genome survey of two-way interacting loci acting within the gametophyte or zygote using independence tests of marker segregations in an F(2) population from an intersubspecific cross between O. sativa subspecies indica and japonica. We detected only one reproducible interaction, and identified paralogous hybrid incompatibility genes, DOPPELGANGER1 (DPL1) and DOPPELGANGER2 (DPL2), by positional cloning. Independent disruptions of DPL1 and DPL2 occurred in indica and japonica, respectively. DPLs encode highly conserved, plant-specific small proteins (â¼10 kDa) and are highly expressed in mature anther. Pollen carrying two defective DPL alleles became nonfunctional and did not germinate, suggesting an essential role for DPLs in pollen germination. Although rice has many duplicated genes resulting from ancient whole genome duplication, the origin of this gene duplication was in recent small-scale gene duplication, occurring after Oryza-Brachypodium differentiation. Comparative analyses suggested the geographic and phylogenetic distribution of these two defective alleles, showing that loss-of-function mutations of DPL1 genes emerged multiple times in indica and its wild ancestor, O. rufipogon, and that the DPL2 gene defect is specific to japonica cultivars.
Assuntos
Genes Duplicados/genética , Genes de Plantas/genética , Especiação Genética , Hibridização Genética , Oryza/genética , Pólen/genética , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar/genética , Componentes do Gene , Frequência do Gene , Teste de Complementação Genética , Genômica/métodos , Germinação/genética , Immunoblotting , Hibridização In Situ , Dados de Sequência Molecular , Mutação/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
Soybean (Glycine max) is one of the most important crop plants for seed protein and oil content, and for its capacity to fix atmospheric nitrogen through symbioses with soil-borne microorganisms. We sequenced the 1.1-gigabase genome by a whole-genome shotgun approach and integrated it with physical and high-density genetic maps to create a chromosome-scale draft sequence assembly. We predict 46,430 protein-coding genes, 70% more than Arabidopsis and similar to the poplar genome which, like soybean, is an ancient polyploid (palaeopolyploid). About 78% of the predicted genes occur in chromosome ends, which comprise less than one-half of the genome but account for nearly all of the genetic recombination. Genome duplications occurred at approximately 59 and 13 million years ago, resulting in a highly duplicated genome with nearly 75% of the genes present in multiple copies. The two duplication events were followed by gene diversification and loss, and numerous chromosome rearrangements. An accurate soybean genome sequence will facilitate the identification of the genetic basis of many soybean traits, and accelerate the creation of improved soybean varieties.
Assuntos
Genoma de Planta/genética , Genômica , Glycine max/genética , Poliploidia , Arabidopsis/genética , Cruzamento , Cromossomos de Plantas/genética , Evolução Molecular , Duplicação Gênica , Genes Duplicados/genética , Genes de Plantas/genética , Dados de Sequência Molecular , Família Multigênica/genética , Filogenia , Nodulação/genética , Locos de Características Quantitativas/genética , Recombinação Genética , Sequências Repetitivas de Ácido Nucleico/genética , Óleo de Soja/biossíntese , Sintenia/genética , Fatores de Transcrição/genéticaRESUMO
The cellular retinoic acid-binding protein type I (CRABPI) is encoded by a single gene in mammals. We have characterized two crabp1 genes in zebrafish, designated crabp1a and crabp1b. These two crabp1 genes share the same gene structure as the mammalian CRABP1 genes and encode proteins that show the highest amino acid sequence identity to mammalian CRABPIs. The zebrafish crabp1a and crabp1b were assigned to linkage groups 25 and 7, respectively. Both linkage groups show conserved syntenies to a segment of the human chromosome 15 harboring the CRABP1 locus. Phylogenetic analysis suggests that the zebrafish crabp1a and crabp1b are orthologs of the mammalian CRABP1 genes that likely arose from a teleost fish lineage-specific genome duplication. Embryonic whole mount in situ hybridization detected zebrafish crabp1b transcripts in the posterior hindbrain and spinal cord from early stages of embryogenesis. crabp1a mRNA was detected in the forebrain and midbrain at later developmental stages. In adult zebrafish, crabp1a mRNA was localized to the optic tectum, whereas crabp1b mRNA was detected in several tissues by RT-PCR but not by tissue section in situ hybridization. The differential and complementary expression patterns of the zebrafish crabp1a and crabp1b genes imply that subfunctionalization may be the mechanism for the retention of both crabp1 duplicated genes in the zebrafish genome.
Assuntos
Duplicação Gênica , Regulação da Expressão Gênica no Desenvolvimento/genética , Genes Duplicados/genética , Genoma , Receptores do Ácido Retinoico/genética , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/genética , Sequência de Aminoácidos , Animais , Cromossomos/genética , DNA Complementar/genética , Humanos , Larva/genética , Larva/crescimento & desenvolvimento , Dados de Sequência Molecular , Especificidade de Órgãos , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores do Ácido Retinoico/química , Alinhamento de Sequência , Transcrição Gênica/genética , Peixe-Zebra/embriologia , Peixe-Zebra/crescimento & desenvolvimento , Proteínas de Peixe-Zebra/químicaRESUMO
INTRODUCTION: Array comparative genomic hybridisation (array CGH) is a powerful method that detects alteration of gene copy number with greater resolution and efficiency than traditional methods. However, its ability to detect disease causing duplications in constitutional genomic DNA has not been shown. We developed an array CGH assay for X linked hypopituitarism, which is associated with duplication of Xq26-q27. METHODS: We generated custom BAC/PAC arrays that spanned the 7.3 Mb critical region at Xq26.1-q27.3, and used them to search for duplications in three previously uncharacterised families with X linked hypopituitarism. RESULTS: Validation experiments clearly identified Xq26-q27 duplications that we had previously mapped by fluorescence in situ hybridisation. Array CGH analysis of novel XH families identified three different Xq26-q27 duplications, which together refine the critical region to a 3.9 Mb interval at Xq27.2-q27.3. Expression analysis of six orthologous mouse genes from this region revealed that the transcription factor Sox3 is expressed at 11.5 and 12.5 days after conception in the infundibulum of the developing pituitary and the presumptive hypothalamus. DISCUSSION: Array CGH is a robust and sensitive method for identifying X chromosome duplications. The existence of different, overlapping Xq duplications in five kindreds indicates that X linked hypopituitarism is caused by increased gene dosage. Interestingly, all X linked hypopituitarism duplications contain SOX3. As mutation of this gene in human beings and mice results in hypopituitarism, we hypothesise that increased dosage of Sox3 causes perturbation of pituitary and hypothalamic development and may be the causative mechanism for X linked hypopituitarism.
Assuntos
Cromossomos Humanos X/genética , Proteínas de Ligação a DNA/genética , Duplicação Gênica , Genes Duplicados/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Proteínas de Grupo de Alta Mobilidade/genética , Hipopituitarismo/genética , Fatores de Transcrição/genética , Adolescente , Adulto , Animais , Criança , Pré-Escolar , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Ligação Genética/genética , Genoma Humano , Humanos , Hipotálamo/embriologia , Hipotálamo/metabolismo , Hibridização in Situ Fluorescente , Lactente , Recém-Nascido , Masculino , Camundongos , Hibridização de Ácido Nucleico , Linhagem , Hipófise/embriologia , Hipófise/metabolismo , Reprodutibilidade dos Testes , Fatores de Transcrição SOXB1RESUMO
Mammalian genomes carry hundreds of Krüppel-type zinc finger (ZNF) genes, most of which reside in familial clusters. ZNF genes encoding Krüppel-associated box (KRAB) motifs are especially prone to this type of tandem organization. Despite their prevalence, little is known about the functions or evolutionary histories of these clustered gene families. Here we describe a homologous pair of human and mouse KRAB-ZNF gene clusters containing 21 human and 10 mouse genes, respectively. Evolutionary analysis uncovered only three pairs of putative orthologs and two cases where a single gene in one species is related to multiple genes in the other; several human genes have no obvious homolog in mouse. We deduce that duplication and loss of ancestral cluster members occurred independently in the primate and rodent lineages after divergence, yielding substantially different ZNF gene repertoires in humans and mice. Differences in expression patterns and sequence divergence within the DNA binding regions of predicted proteins suggest that the duplicated genes have acquired novel functions over evolutionary time. Since KRAB-ZNF proteins are predicted to function as transcriptional regulators, the elaboration of new lineage-specific genes in this and other clustered ZNF families is likely to have had a significant impact on species-specific aspects of biology.
Assuntos
Marcadores Genéticos/genética , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/genética , Dedos de Zinco/genética , Sequência de Aminoácidos , Animais , DNA Complementar/genética , Evolução Molecular , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Genes Duplicados/genética , Humanos , Camundongos , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Estrutura Terciária de Proteína/genética , Seleção Genética , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
The serum amyloid P component (SAP)-like pentraxin Limulus polyphemus SAP is a recently discovered, distinct pentraxin species, of known structure, which does not bind phosphocholine and whose N-terminal sequence has been shown to differ markedly from the highly conserved N terminus of all other known horseshoe crab pentraxins. The complete cDNA sequence of Limulus SAP, and the derived amino acid sequence, the first invertebrate SAP-like pentraxin sequence, have been determined. Two sequences were identified that differed only in the length of the 3' untranslated region. Limulus SAP is synthesised as a precursor protein of 234 amino acid residues, the first 17 residues encoding a signal peptide that is absent from the mature protein. Phylogenetic analysis clusters Limulus SAP pentraxin with the horseshoe crab C-reactive proteins (CRPs) rather than the mammalian SAPs, which are clustered with mammalian CRPs. The deduced amino acid sequence shares 22% identity with both human SAP and CRP, which are 51% identical, and 31-35% with horseshoe crab CRPs. These analyses indicate that gene duplication of CRP (or SAP), followed by sequence divergence and the evolution of CRP and/or SAP function, occurred independently along the chordate and arthropod evolutionary lines rather than in a common ancestor. They further indicate that the CRP/SAP gene duplication event in Limulus occurred before both the emergence of the Limulus CRP variants and the mammalian CRP/SAP gene duplication. Limulus SAP, which does not exhibit the CRP characteristic of calcium-dependent binding to phosphocholine, is established as a pentraxin species distinct from all other known horseshoe crab pentraxins that exist in many variant forms sharing a high level of sequence homology.
Assuntos
Evolução Molecular , Caranguejos Ferradura/genética , Componente Amiloide P Sérico/química , Componente Amiloide P Sérico/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Genes Duplicados/genética , Caranguejos Ferradura/química , Humanos , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Peptídeos/genética , Filogenia , Alinhamento de Sequência , Componente Amiloide P Sérico/isolamento & purificação , Componente Amiloide P Sérico/metabolismoRESUMO
The narrow sheath mutant of maize displays a leaf and plant stature phenotype controlled by the duplicate factor mutations narrow sheath1 and narrow sheath2. Mutant leaves fail to develop a lateral domain that includes the leaf margins. Genetic data are presented to show that the narrow sheath mutations map to duplicated chromosomal regions, reflecting an ancestral duplication of the maize genome. Genetic and cytogenetic evidence indicates that the original mutation at narrow sheath2 is associated with a chromosomal inversion on the long arm of chromosome 4. Meristematic sectors of dual aneuploidy were generated, producing plants genetically mosaic for NARROW SHEATH function. These mosaic plants exhibited characteristic half-plant phenotypes, in which leaves from one side of the plant were of nonmutant morphology and leaves from the opposite side were of narrow sheath mutant phenotype. The data suggest that the narrow sheath duplicate genes may perform ancestrally conserved, redundant functions in the development of a lateral domain in the maize leaf.
Assuntos
Aneuploidia , Sequência Conservada/genética , Genes Duplicados/genética , Folhas de Planta/genética , Zea mays/genética , Alelos , Mapeamento Cromossômico , DNA de Plantas/análise , DNA de Plantas/efeitos da radiação , Mosaicismo/genética , Mutagênese , Fenótipo , Folhas de Planta/química , Folhas de Planta/crescimento & desenvolvimento , Pólen/citologia , Pólen/genética , Polimorfismo de Fragmento de Restrição , Recombinação Genética , Sementes/genética , Sementes/efeitos da radiação , Translocação Genética , Raios X , Zea mays/química , Zea mays/crescimento & desenvolvimentoRESUMO
Two copies of the chloroplast-like tRNA(Asn) gene, trnN1 and trnN2, are expressed in potato mitochondria. While Northern-blot analysis revealed only mature tRNA(Asn), RT-PCR indicated that trnN1 is co-transcribed with trnY and nad2 (exons c, d and e). Using primer-extension and capping experiments, four transcription initiation sites have been mapped in the vicinity of these genes. The first site, responsible for the co-transcription of trnN1, trnY and nad2 (exons c, d and e), gives rise to a primary transcript of at least 7000 nt. A second site, 58 nt downstream from trnY, corresponds to an alternative promoter specific for nad2. In both cases, only the CRTA core motif of the consensus CRTAaGaGA of dicot mitochondrial promoters was found. Finally, two transcription initiation sites were identified 135 and 128 nt upstream of trnN2 in a region which shows no sequence homology with this consensus motif.