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1.
J Insect Physiol ; 121: 104012, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31911184

RESUMO

Pheromone binding proteins (PBPs) are well studied in lepidopteran moths and are considered to be crucial in detection of sex pheromones as well as some green leaf volatiles. In contrast, evidence that PBPs interact with sex pheromones of hemipteran species is not available. The mirid plant bug, Adelphocoris lineolatus (Goeze), is a notorious hemipteran pest that uses two butyrate esters, trans-2-hexenyl butyrate (E2HB) and hexyl butyrate (HB), and one hexenoic aldehyde trans-4-oxo-2-hexenal (E4O2H), as sex pheromones. In the present study, we report on an odorant binding protein, AlinOBP4, with particular focus on its potential physiological roles in the detection of A. lineolatus sex pheromone components. Phylogenetic analyses indicated that AlinOBP4 and two mirid orthologs clustered in a general phylogenetic clade with the lepidopteran ABX OBPs, the fly LUSH and the OBP83a/b subfamily. Cellular localization by fluorescence in situ hybridization and immunolabeling further demonstrated that AlinOBP4 was strongly expressed in the multiporous sensilla trichodea (str) and middle long sensilla basiconica (mlsba) of male A. lineolatus adults, suggesting a key role associated with sex pheromone and odorant detection. A ligand binding assay revealed that recombinant AlinOBP4 protein highly bound not only to the sex pheromone components E4O2H but also to some host plant volatiles. These findings together with the evidence of insect PBPs available in the literature support the view that AlinOBP4 is involved in sex pheromone detection in male A. lineolatus and provide foundational information for further elucidating the molecular mechanisms of chemosensory based mating behavior in hemipteran mirid bugs.

2.
Micron ; 129: 102777, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31811977

RESUMO

Sensilla on antennae of the workers and soldiers of Coptotermes formosanus Shiraki were examined by scanning electron microscopy in this study. As the two castes were allocated totally different tasks in the termite colony, we wondered if there was a big difference between their antennae which were recognized as the main sensory appendages of insects. Therefore, detailed information about the morphology, distribution and abundance of various types of sensilla was described in this report. However, our results showed no obvious caste dimorphism was observed. The morphology of antennae and sensilla as well as the general sensilla distribution pattern did not differ between the workers and soldiers of C. formosanus. In total, seven types of sensilla including sensilla chaetica (I, II, III), Böhm bristles, sensilla campaniformia (I, II, III), sensilla trichodea, sensilla basiconica, sensilla trichodea curvata and sensilla capitula were found on the antennae. Additionally, small apertures were found scattered randomly in the antennal cuticle. Functions of these sensilla or structures were proposed to be mechanoreceptors, chemo-receptors, thermo-hygroreceptors, co2-receptors etc. which probably play crucial roles in their various social behaviors.

3.
Mol Biol Rep ; 47(1): 477-488, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31664595

RESUMO

In this study, two laccase isoenzymes (Lac1 and Lac2) from the culture supernatant of Trametes hirsuta MX2 were purified, and the genes (Lac1 and Lac2) coding the isoenzymes were cloned. Both Lac1 and Lac2 contained an open reading frame of 1563 bp with an identity of 79%. The two isoenzymes showed significant biochemical differences. The maximal activities of Lac1 and Lac2 were at pH 2.5 with 2-2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS), and the optimal temperatures for the activities of Lac1 and Lac2 were 60 and 50 °C, respectively. Lac1 exhibited excellent resistance to acidic conditions and retained 62.17% of its initial activity at pH 2.5 after a 72-h incubation. Lac2 was more thermostable than Lac1 with half-lives (t1/2) of 9.58 and 3.12 h at 50 and 60 °C, respectively; the t1/2 of Lac1 were only 4.19 and 0.88 h, respectively. Both Lac1 and Lac2 isoenzymes have a strong tolerance to Mg2+, Mn2+, Cu2+, and EDTA (50 mM). At a low concentration of 0.05 U mL-1, the enzymes could decolorize towards Remazol Brilliant Blue R, Acid Red 1, Crystal Violet, and Neutral Red in the presence of ABTS. These unusual properties demonstrated that the two laccases have strong potential for specific industrial applications.

4.
J Econ Entomol ; 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31773129

RESUMO

Hermetia illucens L. (the black soldier fly) has received increased attention because of its great potential in converting organic waste into a renewable resource. The prepupae have high proportions of proteins and fats and can serve as feedstuff for livestock and as feedstock for biodiesel production. With the goal to upgrade the conversion of low-value organic wastes into high-value proteins and fat on a large scale, the effects of the feedstuffs food waste, pig manure, chicken manure, and cow dung on the reproductive potential and nutrient composition of H. illucens were evaluated. The intrinsic rate of increase of H. illucens fed food waste (0.1249 d-1) was significantly greater than the rate of those fed pig manure (0.1167 d-1), chicken manure (0.1154 d-1), and cow dung (0.1049 d-1). The ash content of H. illucens fed food waste (30.8 g·kg-1 lyophilized prepupa matter (LPM)) was significantly lower than that of those fed chicken manure (37.6 g·kg-1 LPM) and cow dung (49.5 g·kg-1 LPM). The contents of crude fat, 372.4 g·kg-1 LPM, and protein, 436.9 g·kg-1 LPM, in prepupae fed food waste were the highest among the four treatments. The reproductive performance and prepupal nutrient composition indicated that food waste was the most suitable feed for H. illucens. The results from this study further demonstrate that the prepupae of H. illucens have great potential for use as a protein and fat source in animal feeds and as biodiesel material.

5.
Front Physiol ; 10: 913, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31404332

RESUMO

The functions of cuticular hydrocarbons (CHCs) are varied in insects, but one example is to reduce water loss. Previous work has suggested that biosynthesis of CHCs is strongly related to the CYP4G sub-family. Targeting these genes in the brown planthopper, Nilaparvata lugens Stål, might be a new application for integrated pest management. Therefore, we explored the functions of CYP4G76 (GenBank: KM217045.1) and CYP4G115 (GenBank: KM217046.1) genes in this study. The desiccation treatment (RH < 5%) for the duration of 1-3 days significantly increased the transcription level of CYP4G76 and CYP4G115. RNAi through the injection of CYP4G76 and CYP4G115 dsRNA could significantly decrease their expression, respectively, and further reduced the biosynthesis of CHCs, i.e., saturated and straight-chain alkanes. When CYP4G76 and CYP4G115 were suppressed, the susceptibility of N. lugens nymphs to desiccation increased, due to the deficiency of the CHCs in the insect's cuticle. When the expression of CYP4G76 and CYP4G115 was decreased, this resulted in an increased rate of penetration of the four insecticides: pymetrozine, imidacloprid, thiamethoxam and buprofezin. Therefore, CYP4G76 and CYP4G115 appear to regulate the biosynthesis of CHCs in N. lugens nymphs, which play a major role in protecting insects from water loss and the penetration of insecticides. CYP4G76 and CYP4G115 might be used as a novel target in integrated pest management to N. lugens.

6.
Environ Sci Pollut Res Int ; 26(23): 23824-23831, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31209751

RESUMO

The toxicity of Origanum vulgare essential oil to the housefly Musca domestica L. was evaluated. The major constituents of the O. vulgare essential oil by gas chromatographic mass spectrometry (GC-MS) analysis were carvacrol (58.13%), p-cymene (17.85%), thymol (8.15%), γ-terpinene (4.96%), and linalool (3.69%). Toxicity of O. vulgare essential oil against larvae and pupae was evaluated using fumigation and contact assays. The contact toxicity (LC50) of O. vulgare essential oil and carvacrol for larvae was 0.23 and 0.03 µL/cm2, respectively. The fumigation toxicity (LC50) of O. vulgare essential oil and carvacrol for larvae was 9.52 and 2.78 µL/L, respectively. Pupal toxicity was evaluated by percentage inhibition rate (PIR). PIR of O. vulgare essential oil at 0.25 µL/cm2 was 90.9% for the contact assay and 100% at 20 µL/L for the fumigation assay. PIR of carvacrol was 29.5% (0.025 µL/cm2) and 81.8% (1.25 µL/L) for the contact toxicity and fumigation assay, respectively. O. vulgare essential oil and carvacrol have significant toxicity to the housefly and are potential insecticides for housefly control.


Assuntos
Moscas Domésticas/efeitos dos fármacos , Inseticidas/análise , Larva/efeitos dos fármacos , Óleos Voláteis/química , Origanum/química , Pupa/efeitos dos fármacos , Timol/análise , Animais , Fumigação , Cromatografia Gasosa-Espectrometria de Massas , Monoterpenos/química , Muscidae/química
8.
Nanotechnology ; 29(18): 185602, 2018 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-29451119

RESUMO

The interfacial microcracks in the resin matrix composites are difficult to be detected and repaired. However, the self-healing concept provides opportunities to fabricate composites with unusual properties. In the present study, photothermal conversion Ag-Cu2S nanoparticles were immobilized onto poly(p-phenylene benzobisoxazole) (PBO) fibers via a polydopamine chemistry. Benefitting from the photothermal effects of Ag-Cu2S, the obtained PBO fibers (Ag-Cu2S-PBO) efficiently converted the light energy into heat under Xenon lamp irradiation. Then, single PBO fiber composites were prepared using thermoplastic polyurethane as the matrix. It was found that the interfacial damage caused by single fiber pull-out was simply self-healed by Xe light irradiation. This wonderful interfacial damage self-healing property was mainly attributed to the in situ heating generation via photothermal effects of Ag-Cu2S in the composite interface. This paper reports a novel strategy to construct advanced composites with light-triggered self-healing properties, which will provide inspiration for preparing high performance composite materials.

9.
Sci Rep ; 7(1): 10990, 2017 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-28887463

RESUMO

The elongases of very long chain fatty acid (ELOVL or ELO) are essential in the biosynthesis of fatty acids longer than C14. Here, two ELO full-length cDNAs (TmELO1, TmELO2) from the yellow mealworm (Tenebrio molitor L.) were isolated and the functions were characterized. The open reading frame (ORF) lengths of TmELO1 and TmELO2 were 1005 bp and 972 bp, respectively and the corresponding peptide sequences each contained several conserved motifs including the histidine-box motif HXXHH. Phylogenetic analysis demonstrated high similarity with the ELO of Tribolium castaneum and Drosophila melanogaster. Both TmELO genes were expressed at various levels in eggs, 1st and 2nd instar larvae, mature larvae, pupae, male and female adults. Injection of dsTmELO1 but not dsTmELO2 RNA into mature larvae significantly increased mortality although RNAi did not produce any obvious changes in the fatty acid composition in the survivors. Heterologous expression of TmELO genes in yeast revealed that TmELO1 and TmELO2 function to synthesize long chain and very long chain fatty acids.


Assuntos
Acetiltransferases/metabolismo , Tenebrio/metabolismo , Acetiltransferases/química , Acetiltransferases/genética , Sequência de Aminoácidos , Animais , Ácidos Graxos/metabolismo , Expressão Gênica , Fases de Leitura Aberta , Filogenia , Interferência de RNA , Tenebrio/classificação , Tenebrio/genética
10.
Materials (Basel) ; 10(6)2017 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-28773033

RESUMO

In this study, the synthesis of the core/shell structured diamond/akageneite hybrid particles was performed through one-step isothermal hydrolyzing. The hybrid particle was characterized by X-ray diffraction, field emission scanning electron microscopy, and Fourier transform infrared spectra. The test results overall reveal that the akageneite coating, phase ß-FeO(OH), was uniformly coated onto the diamond surface. The polishing performance of the pristine diamond and hybrid particles for the sapphire substrate was evaluated respectively. The experimental results show that the hybrid particles exhibited improved polishing quality and prolonged effective processing time of polishing pad compared with diamond particles without compromising the material remove rate and surface roughness. The improved polishing behavior might be attributed to the ß-FeOOH coating, which is conducive to less abrasive shedding and reducing the scratch depth.

11.
Nanoscale ; 9(25): 8825-8833, 2017 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-28632263

RESUMO

The wonderful chemical structures and characteristics of low-dimensional carbon materials have exciting applications in life sciences. In the present study, we developed a facile strategy to conjugate C60 with graphene via host-guest chemistry for targeted phototherapy. A versatile carrier based on folic acid-functionalized graphene (GO-FA) and comprising γ-cyclodextrin (γ-CD) at its surface was assembled via π-π interaction, creating hybrid structures with drug storage and tumor targeting properties. This γ-CD-modified graphene (GO-FA/Py-γ-CD) is capable of hosting pristine C60 molecules for the fabrication of a GO-FA/Py-γ-CD/C60 nanohybrid. The hybridization of GO-FA, γ-CD, and C60 hinders the aggregation of C60, promotes cellular uptake, enhances light absorption, and finally demonstrates enhanced phototherapy effects of GO-FA/Py-γ-CD/C60. Under Xe lamp irradiation (2 W cm-2) for 4 min, GO-FA/Py-γ-CD/C60 simultaneously causes heating and intracellular ROS production, which further significantly reduces the cell viability to 16.2% at low content of loading (30 µg mL-1). Moreover, it represents an excellent tumor killing efficiency, better than that of the other reported graphene/C60 nanohybrids; thus, this material is suitable for applications in phototherapy of cancer.


Assuntos
Portadores de Fármacos , Fulerenos , Grafite , Nanopartículas , Fotoquimioterapia , gama-Ciclodextrinas , Sobrevivência Celular , Ácido Fólico , Células HeLa , Humanos , Neoplasias/tratamento farmacológico
12.
Environ Entomol ; 46(4): 916-925, 2017 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-28535309

RESUMO

Mating disruption of Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae) with its sex pheromone has not been commonly used in NE China due to a lack of information about optimal sex pheromone dosages and the density of release points required in the field. During 2014-2016, first, the two active pheromone ingredients were evaluated in the laboratory alone at ca. 2.5-5.0 mg, or in combination at 0.2-6.0 mg, to disrupt male O. furnacalis mating behaviors. Then, mating disruption areas, with radii of <8.0 m, were determined with those same dosages in corn, an orchard, and soybean fields by comparing male captures in sentinel traps in the control plots with those in corresponding disruption treatments. Finally, 6.0 (F30) and 0.2 mg (Fs) dosages were used in fields at 20-640 and 200-6,400 release points/ha. We found that ≧6.0 mg of the binary pheromone mixture, or ca. 5.0 mg of either of the two single components, completely disrupted mating behaviors, and F30 of the binary mixture provided a 200-m2 disruption area, with at least 50% capture reductions. At a density of 60-640 and 600-6,400 points/ha in a corn field, F30 and Fs dosages provided >90% mating disruption, leaf protection, and ear protection. The dispenser densities and inverse male catches in traps tended to follow a noncompetitive mechanism of mating disruption. Since 85% disruption of mating with 200-400 0.02 mg release points/ha was obtained, that level is recommended as the choice in future NE China O. furnacalis IPM programs.


Assuntos
Controle de Insetos , Atrativos Sexuais/farmacologia , Comportamento Sexual Animal/efeitos dos fármacos , Animais , China , Relação Dose-Resposta a Droga , Masculino , Zea mays/crescimento & desenvolvimento
13.
J Insect Sci ; 16(1)2016.
Artigo em Inglês | MEDLINE | ID: mdl-27271970

RESUMO

Fatty acyl-CoA reductases (FARs) are key enzymes involved in fatty alcohol synthesis. Here, we cloned and characterized full-length cDNAs of two FAR genes from the cotton mealybug, Phenacoccus solenopsis. The results showed PsFAR I and PsFAR II cDNAs were 1,584 bp and 1,515 bp in length respectively. Both PsFAR I and PsFAR II were predicted to be located in the endoplasmic reticulum by Euk-mPLoc 2.0 approach. Both of them had a Rossmann folding region and a FAR_C region. Two conservative motifs were discovered in Rossmann folding region by sequence alignment including a NADPH combining motif, TGXXGG, and an active site motif, YXXXK. A phylogenetic tree made using MEGA 6.06 indicated that PsFAR I and PsFAR II were placed in two different branches. Gene expression analysis performed at different developmental stages showed that the expression of PsFar I is significantly higher than that of PsFar II in first and second instar nymphs and in male adults. Spirotetramat treatment at 125 mg/liter significantly increased the expression of PsFar I in third instar nymphs, but there was no effect in the expression of PsFar II Our results indicated these two FAR genes showed different expression patterns during insect development and after pesticide treatment, suggesting they play different roles in insect development and detoxification against pesticides.


Assuntos
Aldeído Oxirredutases/genética , Hemípteros/enzimologia , Hemípteros/genética , Proteínas de Insetos/genética , Aldeído Oxirredutases/química , Aldeído Oxirredutases/metabolismo , Sequência de Aminoácidos , Animais , Hemípteros/classificação , Hemípteros/crescimento & desenvolvimento , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Masculino , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Ninfa/metabolismo , Filogenia , Alinhamento de Sequência
14.
Genetics ; 201(4): 1307-18, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26450921

RESUMO

We describe an adaptation of φC31 integrase-mediated targeted cassette exchange for use in Drosophila cell lines. Single copies of an attP-bounded docking platform carrying a GFP-expression marker, with or without insulator elements flanking the attP sites, were inserted by P-element transformation into the Kc167 and Sg4 cell lines; each of the resulting docking-site lines carries a single mapped copy of one of the docking platforms. Vectors for targeted substitution contain a cloning cassette flanked by attB sites. Targeted substitution occurs by integrase-mediated substitution between the attP sites (integrated) and the attB sites (vector). We describe procedures for isolating cells carrying the substitutions and for eliminating the products of secondary off-target events. We demonstrate the technology by integrating a cassette containing a Cu(2+)-inducible mCherry marker, and we report the expression properties of those lines. When compared with clonal lines made by traditional transformation methods, which lead to the illegitimate insertion of tandem arrays, targeted insertion lines give more uniform expression, lower basal expression, and higher induction ratios. Targeted substitution, though intricate, affords results that should greatly improve comparative expression assays-a major emphasis of cell-based studies.


Assuntos
Drosophila melanogaster/genética , Marcação de Genes/métodos , Genoma de Inseto , Mutagênese Insercional , Animais , Linhagem Celular , Células Clonais , Engenharia Genética , Marcadores Genéticos , Integrases/metabolismo , Transgenes
15.
J Econ Entomol ; 108(2): 701-6, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26470181

RESUMO

The foraging activities, including foraging range and seasonal fluctuation of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were investigated in subtropical areas in China. Six mature C. formosanus colonies were selected for this study. Foraging distance and area were conducted in Anqing1, Hengyang, and Wuxi1 colonies, while the seasonal fluctuation was conducted in Wuxi2, Wuwei, and Anqing2 colonies. Mark-release-recapture method analysis showed that the Formosan termites foraged at least 11.5, 28.7, and 56.8 m away from the main nest and covered 98.1, 543.7, and 671.9 m2 of foraging area at Anqing1, Hengyang, and Wuxi1 site, respectively. The seasonal fluctuation in termite colony activity showed an "M shape" pattern according to the wood damages caused by termites at monitoring stations. Peak colony activity at Wuxi2, Wuwei, and Anqing2 occurred in July and October, June and September, July and October, respectively. This study provides critical information for the integrated management of C. formosanus, including baiting application in the subtropical regions of China, where it constitutes the most destructive pest for household structures.


Assuntos
Comportamento Animal , Isópteros , Animais , China , Madeira
16.
PLoS One ; 10(3): e0121445, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25803778

RESUMO

ß-Galactosidases can transfer the galactosyl from lactose or galactoside donors to various acceptors and thus are especially useful for the synthesis of important glycosides. However, these enzymes have limitations in the glycosylation of phenolic compounds that have many physiological functions. In this work, the ß-galactosidase from Lactobacillus bulgaricus L3 was subjected to site-saturation mutagenesis at the W980 residue. The recombinant pET-21b plasmid carrying the enzyme gene was used as the template for mutation. The mutant plasmids were transformed into Escherichia coli cells for screening. One recombinant mutant, W980F, exhibited increased yield of glycoside when using hydroquinone as the screening acceptor. The enzyme was purified and the effects of the mutation on enzyme properties were determined in detail. It showed improved transglycosylation activity on novel phenolic acceptors besides hydroquinone. The yields of the glycosides produced from phenol, hydroquinone, and catechol were increased by 7.6% to 53.1%. Moreover, it generated 32.3% glycosides from the pyrogallol that could not be glycosylated by the wild-type enzyme. Chemical structures of these glycoside products were further determined by MS and NMR analysis. Thus, a series of novel phenolic galactosides were achieved by ß-galactosidase for the first time. This was a breakthrough in the enzymatic galactosylation of the challenging phenolic compounds of great values.


Assuntos
Lactobacillus/enzimologia , Mutagênese Sítio-Dirigida , Mutação , Fenóis/metabolismo , beta-Galactosidase/genética , beta-Galactosidase/metabolismo , Sequência de Aminoácidos , Glicosilação , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Especificidade por Substrato , beta-Galactosidase/química
17.
Genome Biol ; 15(8): R70, 2014 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-25262759

RESUMO

BACKGROUND: Structural rearrangements of the genome resulting in genic imbalance due to copy number change are often deleterious at the organismal level, but are common in immortalized cell lines and tumors, where they may be an advantage to cells. In order to explore the biological consequences of copy number changes in the Drosophila genome, we resequenced the genomes of 19 tissue-culture cell lines and generated RNA-Seq profiles. RESULTS: Our work revealed dramatic duplications and deletions in all cell lines. We found three lines of evidence indicating that copy number changes were due to selection during tissue culture. First, we found that copy numbers correlated to maintain stoichiometric balance in protein complexes and biochemical pathways, consistent with the gene balance hypothesis. Second, while most copy number changes were cell line-specific, we identified some copy number changes shared by many of the independent cell lines. These included dramatic recurrence of increased copy number of the PDGF/VEGF receptor, which is also over-expressed in many cancer cells, and of bantam, an anti-apoptosis miRNA. Third, even when copy number changes seemed distinct between lines, there was strong evidence that they supported a common phenotypic outcome. For example, we found that proto-oncogenes were over-represented in one cell line (S2-DRSC), whereas tumor suppressor genes were under-represented in another (Kc167). CONCLUSION: Our study illustrates how genome structure changes may contribute to selection of cell lines in vitro. This has implications for other cell-level natural selection progressions, including tumorigenesis.


Assuntos
Linhagem Celular , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Evolução Molecular , Dosagem de Genes , Animais , Sobrevivência Celular , DNA/análise , Proteínas de Drosophila/genética , Feminino , Aptidão Genética , Variação Genética , Masculino , MicroRNAs/genética , Receptores Proteína Tirosina Quinases/genética , Seleção Genética , Análise de Sequência de DNA , Cromossomos Sexuais/genética , Técnicas de Cultura de Tecidos
18.
Genome Res ; 24(7): 1236-50, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24985917

RESUMO

We expanded the knowledge base for Drosophila cell line transcriptomes by deeply sequencing their small RNAs. In total, we analyzed more than 1 billion raw reads from 53 libraries across 25 cell lines. We verify reproducibility of biological replicate data sets, determine common and distinct aspects of miRNA expression across cell lines, and infer the global impact of miRNAs on cell line transcriptomes. We next characterize their commonalities and differences in endo-siRNA populations. Interestingly, most cell lines exhibit enhanced TE-siRNA production relative to tissues, suggesting this as a common aspect of cell immortalization. We also broadly extend annotations of cis-NAT-siRNA loci, identifying ones with common expression across diverse cells and tissues, as well as cell-restricted loci. Finally, we characterize small RNAs in a set of ovary-derived cell lines, including somatic cells (OSS and OSC) and a mixed germline/somatic cell population (fGS/OSS) that exhibits ping-pong piRNA signatures. Collectively, the ovary data reveal new genic piRNA loci, including unusual configurations of piRNA-generating regions. Together with the companion analysis of mRNAs described in a previous study, these small RNA data provide comprehensive information on the transcriptional landscape of diverse Drosophila cell lines. These data should encourage broader usage of fly cell lines, beyond the few that are presently in common usage.


Assuntos
Drosophila/genética , Variação Genética , MicroRNAs/genética , RNA Interferente Pequeno/genética , Animais , Sequência de Bases , Linhagem Celular , Biologia Computacional/métodos , Expressão Gênica , Loci Gênicos , Células Germinativas , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/química , Anotação de Sequência Molecular , Dados de Sequência Molecular , Conformação de Ácido Nucleico , RNA Interferente Pequeno/química , Alinhamento de Sequência
19.
Nature ; 512(7515): 393-9, 2014 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-24670639

RESUMO

Animal transcriptomes are dynamic, with each cell type, tissue and organ system expressing an ensemble of transcript isoforms that give rise to substantial diversity. Here we have identified new genes, transcripts and proteins using poly(A)+ RNA sequencing from Drosophila melanogaster in cultured cell lines, dissected organ systems and under environmental perturbations. We found that a small set of mostly neural-specific genes has the potential to encode thousands of transcripts each through extensive alternative promoter usage and RNA splicing. The magnitudes of splicing changes are larger between tissues than between developmental stages, and most sex-specific splicing is gonad-specific. Gonads express hundreds of previously unknown coding and long non-coding RNAs (lncRNAs), some of which are antisense to protein-coding genes and produce short regulatory RNAs. Furthermore, previously identified pervasive intergenic transcription occurs primarily within newly identified introns. The fly transcriptome is substantially more complex than previously recognized, with this complexity arising from combinatorial usage of promoters, splice sites and polyadenylation sites.


Assuntos
Drosophila melanogaster/genética , Perfilação da Expressão Gênica , Transcriptoma/genética , Processamento Alternativo/genética , Animais , Drosophila melanogaster/anatomia & histologia , Drosophila melanogaster/citologia , Feminino , Masculino , Anotação de Sequência Molecular , Tecido Nervoso/metabolismo , Especificidade de Órgãos , Poli A/genética , Poliadenilação , Regiões Promotoras Genéticas/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Caracteres Sexuais , Estresse Fisiológico/genética
20.
Cell Rep ; 1(3): 277-89, 2012 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-22685694

RESUMO

We analyzed the usage and consequences of alternative cleavage and polyadenylation (APA) in Drosophila melanogaster by using >1 billion reads of stranded mRNA-seq across a variety of dissected tissues. Beyond demonstrating that a majority of fly transcripts are subject to APA, we observed broad trends for 3' untranslated region (UTR) shortening in the testis and lengthening in the central nervous system (CNS); the latter included hundreds of unannotated extensions ranging up to 18 kb. Extensive northern analyses validated the accumulation of full-length neural extended transcripts, and in situ hybridization indicated their spatial restriction to the CNS. Genes encoding RNA binding proteins (RBPs) and transcription factors were preferentially subject to 3' UTR extensions. Motif analysis indicated enrichment of miRNA and RBP sites in the neural extensions, and their termini were enriched in canonical cis elements that promote cleavage and polyadenylation. Altogether, we reveal broad tissue-specific patterns of APA in Drosophila and transcripts with unprecedented 3' UTR length in the nervous system.


Assuntos
Drosophila melanogaster/genética , Especificidade de Órgãos/genética , Poliadenilação/genética , Regiões 3' não Traduzidas/genética , Animais , Sequência de Bases , Northern Blotting , Sequência Conservada/genética , Proteínas de Ligação a DNA/metabolismo , Drosophila melanogaster/embriologia , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Genes de Insetos/genética , Hibridização In Situ , Masculino , Dados de Sequência Molecular , Neurônios/citologia , Neurônios/metabolismo , Motivos de Nucleotídeos/genética , Poli A/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Análise de Sequência de RNA , Testículo/metabolismo , Transcriptoma/genética
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