RESUMO
Taenia solium cysts were collected from pig skeletal muscle and analyzed via a shotgun proteomic approach to identify known proteins in the cyst fluid and to explore host-parasite interactions. Cyst fluid was aseptically collected and analyzed with shotgun liquid chromatography-tandem mass spectrometry (LC-MS/MS). Gene alignment and annotation were performed using Blast2GO software followed by gene ontology analysis of the annotated proteins. The pathways were further analyzed with the Kyoto Encyclopedia of Genes and Genomes (KEGG), and a protein-protein interaction (PPI) network map was generated using STRING software. A total of 158 known proteins were identified, most of which were low-molecular-mass proteins. These proteins were mainly involved in cellular and metabolic processes, and their molecular functions were predominantly related to catalytic activity and binding functions. The pathway enrichment analysis revealed that the known proteins were mainly enriched in the PI3K-Akt and glycolysis/gluconeogenesis signaling pathways. The nodes in the PPI network mainly consisted of enzymes involved in sugar metabolism. The cyst fluid proteins screened in this study may play important roles in the interaction between the cysticerci and the host. The shotgun LC-MS/MS, gene ontology, KEGG, and PPI network map data will be used to identify and analyze the cyst fluid proteome of cysticerci, which will provide a basis for further exploration of the invasion and activities of T. solium.
Assuntos
Proteínas de Helminto/análise , Proteômica/métodos , Taenia solium/química , Animais , Cromatografia Líquida , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Interações Hospedeiro-Parasita , Anotação de Sequência Molecular/métodos , Peso Molecular , Músculo Esquelético/parasitologia , Mapas de Interação de Proteínas , Alinhamento de Sequência , Transdução de Sinais , Suínos , Taenia solium/genética , Espectrometria de Massas em TandemRESUMO
Parasitic nematodes are highly successful pathogens, inflicting disease on humans, animals and plants. Despite great differences in their life cycles, host preference and transmission modes, these parasites share a common capacity to manipulate their host's immune system. This is at least partly achieved through the release of excretory/secretory proteins, the most well-characterized component of nematode secretomes, that are comprised of functionally diverse molecules. In this work, we analyzed published protein secretomes of parasitic nematodes to identify common patterns as well as species-specific traits. The 20 selected organisms span 4 nematode clades, including plant pathogens, animal parasites, and the free-living species Caenorhabditis elegans. Transthyretin-like proteins were the only component common to all adult secretomes; many other protein classes overlapped across multiple datasets. The glycolytic enzymes aldolase and enolase were present in all parasitic species, but missing from C. elegans. Secretomes from larval stages showed less overlap between species. Although comparison of secretome composition across species and life-cycle stages is challenged by the use of different methods and depths of sequencing among studies, our workflow enabled the identification of conserved protein families and pinpointed elements that may have evolved as to enable parasitism. This strategy, extended to more secretomes, may be exploited to prioritize therapeutic targets in the future.
Assuntos
Proteínas de Helminto/metabolismo , Especificidade de Hospedeiro , Nematoides/fisiologia , Secretoma/metabolismo , Animais , Caenorhabditis elegans , Feminino , Proteínas de Helminto/classificação , Humanos , Estágios do Ciclo de Vida , Masculino , Filogenia , Especificidade da EspécieRESUMO
The rice root-knot nematode Meloidogyne graminicola is a major biotic stress for the rice crop under upland, rain-fed lowland and irrigated cultivation conditions. Here, we present an improved draft genome assembly of M. graminicola IARI strain using the long-read sequencing approach (PacBio Sequel platform). The assembled genome size was 36.86 Mb with 514 contigs and N50 value of 105 kb. BUSCO estimated the genome to be 88.6% complete. Meloidogyne graminicola genome contained 17.83% repeat elements and showed 14,062 protein-coding gene models, 4,974 conserved orthologous genes, 561 putative secreted proteins, 49 RNAi pathway genes, 1,853 proteins involved in pathogen-host interactions, 1,575 carbohydrate-active enzymes, and 32,138 microsatellites. Five of the carbohydrate-active enzymes were found only in M. graminicola genome and were not present in any other analysed root-knot nematode genome. Together with the previous two genome assemblies, this improved genome assembly would facilitate comparative and functional genomics for M. graminicola.
Assuntos
Genes de Helmintos , Genoma Helmíntico , Proteínas de Helminto/genética , Oryza/parasitologia , Tylenchoidea/genética , Animais , Ontologia Genética , Tamanho do Genoma , Proteínas de Helminto/classificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Repetições de Microssatélites , Anotação de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Doenças das Plantas/parasitologia , Tylenchoidea/classificaçãoRESUMO
The ability to respond to light has profoundly shaped life. Animals with eyes overwhelmingly rely on their visual circuits for mediating light-induced coordinated movements. Building on previously reported behaviors, we report the discovery of an organized, eye-independent (extraocular), body-wide photosensory framework that allows even a head-removed animal to move like an intact animal. Despite possessing sensitive cerebral eyes and a centralized brain that controls most behaviors, head-removed planarians show acute, coordinated ultraviolet-A (UV-A) aversive phototaxis. We find this eye-brain-independent phototaxis is mediated by two noncanonical rhabdomeric opsins, the first known function for this newly classified opsin-clade. We uncover a unique array of dual-opsin-expressing photoreceptor cells that line the periphery of animal body, are proximal to a body-wide nerve net, and mediate UV-A phototaxis by engaging multiple modes of locomotion. Unlike embryonically developing cerebral eyes that are functional when animals hatch, the body-wide photosensory array matures postembryonically in "adult-like animals." Notably, apart from head-removed phototaxis, the body-wide, extraocular sensory organization also impacts physiology of intact animals. Low-dose UV-A, but not visible light (ocular-stimulus), is able to arouse intact worms that have naturally cycled to an inactive/rest-like state. This wavelength selective, low-light arousal of resting animals is noncanonical-opsin dependent but eye independent. Our discovery of an autonomous, multifunctional, late-maturing, organized body-wide photosensory system establishes a paradigm in sensory biology and evolution of light sensing.
Assuntos
Encéfalo/metabolismo , Olho/metabolismo , Proteínas de Helminto/genética , Opsinas/genética , Células Fotorreceptoras de Invertebrados/metabolismo , Planárias/genética , Animais , Nível de Alerta/genética , Nível de Alerta/fisiologia , Nível de Alerta/efeitos da radiação , Encéfalo/crescimento & desenvolvimento , Olho/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Hibridização in Situ Fluorescente/métodos , Locomoção/genética , Locomoção/fisiologia , Locomoção/efeitos da radiação , Movimento/fisiologia , Movimento/efeitos da radiação , Opsinas/classificação , Opsinas/metabolismo , Filogenia , Planárias/crescimento & desenvolvimento , Planárias/metabolismo , Interferência de RNA , Raios UltravioletaRESUMO
In the absence of efficient alternative strategies, the control of parasitic nematodes, impacting human and animal health, mainly relies on the use of broad-spectrum anthelmintic compounds. Unfortunately, most of these drugs have a limited single-dose efficacy against infections caused by the whipworm, Trichuris. These infections are of both human and veterinary importance. However, in contrast to a wide range of parasitic nematode species, the narrow-spectrum anthelmintic oxantel has a high efficacy on Trichuris spp. Despite this knowledge, the molecular target(s) of oxantel within Trichuris is still unknown. In the distantly related pig roundworm, Ascaris suum, oxantel has a small, but significant effect on the recombinant homomeric Nicotine-sensitive ionotropic acetylcholine receptor (N-AChR) made up of five ACR-16 subunits. Therefore, we hypothesized that in whipworms, a putative homolog of an ACR-16 subunit, can form a functional oxantel-sensitive receptor. Using the pig whipworm T. suis as a model, we identified and cloned a novel ACR-16-like subunit and successfully expressed the corresponding homomeric channel in Xenopus laevis oocytes. Electrophysiological experiments revealed this receptor to have distinctive pharmacological properties with oxantel acting as a full agonist, hence we refer to the receptor as an O-AChR subtype. Pyrantel activated this novel O-AChR subtype moderately, whereas classic nicotinic agonists surprisingly resulted in only minor responses. We observed that the expression of the ACR-16-like subunit in the free-living nematode Caenorhabditis elegans conferred an increased sensitivity to oxantel of recombinant worms. We demonstrated that the novel Tsu-ACR-16-like receptor is indeed a target for oxantel, although other receptors may be involved. These finding brings new insight into the understanding of the high sensitivity of whipworms to oxantel, and highlights the importance of the discovery of additional distinct receptor subunit types within Trichuris that can be used as screening tools to evaluate the effect of new synthetic or natural anthelmintic compounds.
Assuntos
Antinematódeos/farmacologia , Proteínas de Helminto/antagonistas & inibidores , Pirantel/análogos & derivados , Receptores Colinérgicos/química , Tricuríase/tratamento farmacológico , Trichuris/efeitos dos fármacos , Animais , Caenorhabditis elegans/efeitos dos fármacos , Feminino , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Masculino , Pirantel/farmacologia , Receptores Colinérgicos/classificação , Receptores Colinérgicos/metabolismo , Suínos , Tricuríase/metabolismo , Tricuríase/parasitologia , Xenopus laevis/metabolismoRESUMO
The powerful regenerative ability of planarians has long been a concern of scientists, but recently, their efficient immune system has attracted more and more attention from researchers. Gamma-interferon-inducible lysosomal thiol reductase (GILT) is related not only to antigen presentation but also to bacteria invasions. But the systematic studies are not yet to be conducted on the relationship between bacterial infection. Our study reveals for the first time that GILT of planarian (DjGILT) plays an essential role in the clearance of Gram-negative bacteria by conducting H2O2 concentration in planarians. In animals that DjGILT was silenced, it persisted for up to 9 days before all bacteria were cleared, compared with 6 days of the control group. When infected with E. coli and V. anguillarum, the level of H2O2 was significantly increased in DjGILT-silenced planarians, and concomitantly, mRNA level of C-type lectin DjCTL, which modulates agglutination and clearance efficiency of invading bacteria, was decreased. Further study showed that the decrease of H2O2 level led to a significant increase in DjCTL transcripts. Collectively, we proposed a mechanism model for the involvement of GILT gene in bacterial elimination. We have for the first time revealed the specific mechanism of GILT in innate immune response against bacterial infection.
Assuntos
Bactérias Gram-Negativas/imunologia , Proteínas de Helminto/imunologia , Interferon gama/farmacologia , Lisossomos/efeitos dos fármacos , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/imunologia , Planárias/imunologia , Sequência de Aminoácidos , Animais , Escherichia coli/imunologia , Escherichia coli/fisiologia , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/imunologia , Bactérias Gram-Negativas/fisiologia , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Interações Hospedeiro-Patógeno/imunologia , Peróxido de Hidrogênio/imunologia , Peróxido de Hidrogênio/metabolismo , Imunidade Inata/genética , Imunidade Inata/imunologia , Lisossomos/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/genética , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/metabolismo , Filogenia , Planárias/genética , Planárias/microbiologia , Homologia de Sequência de Aminoácidos , Compostos de Sulfidrila/metabolismo , Vibrio/imunologia , Vibrio/fisiologiaRESUMO
P-glycoproteins (Pgp) have been proposed as contributors to the widespread macrocyclic lactone (ML) resistance in several nematode species including a major pathogen of foals, Parascaris univalens. Using new and available RNA-seq data, ten different genomic loci encoding Pgps were identified and characterized by transcriptome-guided RT-PCRs and Sanger sequencing. Phylogenetic analysis revealed an ascarid-specific Pgp lineage, Pgp-18, as well as two paralogues of Pgp-11 and Pgp-16. Comparative gene expression analyses in P. univalens and Caenorhabditis elegans show that the intestine is the major site of expression but individual gene expression patterns were not conserved between the two nematodes. In P. univalens, PunPgp-9, PunPgp-11.1 and PunPgp-16.2 consistently exhibited the highest expression level in two independent transcriptome data sets. Using RNA-Seq, no significant upregulation of any Pgp was detected following in vitro incubation of adult P. univalens with ivermectin suggesting that drug-induced upregulation is not the mechanism of Pgp-mediated ML resistance. Expression and functional analyses of PunPgp-2 and PunPgp-9 in Saccharomyces cerevisiae provide evidence for an interaction with ketoconazole and ivermectin, but not thiabendazole. Overall, this study established reliable reference gene models with significantly improved annotation for the P. univalens Pgp repertoire and provides a foundation for a better understanding of Pgp-mediated anthelmintic resistance.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Ascaridoidea/genética , Proteínas de Helminto/genética , Cavalos/parasitologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/classificação , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Antiparasitários/farmacologia , Infecções por Ascaridida/tratamento farmacológico , Infecções por Ascaridida/parasitologia , Ascaridoidea/metabolismo , Ascaridoidea/fisiologia , Resistência a Medicamentos/efeitos dos fármacos , Resistência a Medicamentos/genética , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Ivermectina/farmacologia , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA/métodos , Análise de Sequência de RNA/métodos , Análise de Sequência de RNA/estatística & dados numéricos , TranscriptomaRESUMO
The tropical liver fluke, Fasciola gigantica is a food-borne parasite responsible for the hepatobiliary disease fascioliasis. The recent completion of F. gigantica genome sequencing by our group has provided a platform for the systematic analysis of the parasite genome. Eukaryotic protein kinases (ePKs) are regulators of cellular phosphorylation. In the present study, we used various computational and bioinformatics tools to extensively analyse the ePKs in F. gigantica (FgePKs) genome. A total of 455 ePKs were identified that represent ~2% of the parasite genome. Out of these, 214 ePKs are typical kinases (Ser/Thr- and Tyr-specific ePKs), and 241 were other kinases. Several FgePKs were found to possess unusual domain architectures, which suggests the diverse nature of the proteins that can be exploited for designing novel inhibitors. 115 kinases showed <35% query coverage when compared to human ePKs highlighting significant divergences in their respective kinomes, further providing a platform for novel structure-based drug designing. This study provides a platform that may open new avenues into our understanding of helminth biochemistry and drug discovery.
Assuntos
Células Eucarióticas/enzimologia , Fasciola/genética , Genoma Helmíntico/genética , Estudo de Associação Genômica Ampla/métodos , Proteínas de Helminto/genética , Proteínas Quinases/genética , Animais , Biologia Computacional/métodos , Fasciola/enzimologia , Fasciola/fisiologia , Fasciolíase/parasitologia , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Humanos , Isoenzimas/classificação , Isoenzimas/genética , Isoenzimas/metabolismo , Família Multigênica/genética , Fosforilação , Filogenia , Proteínas Quinases/classificação , Proteínas Quinases/metabolismoRESUMO
Protein ADP-ribosylation is a reversible post-translational modification (PTM) process that plays fundamental roles in cell signaling. The covalent attachment of ADP ribose polymers is executed by PAR polymerases (PARP) and it is essential for chromatin organization, DNA repair, cell cycle, transcription, and replication, among other critical cellular events. The process of PARylation or polyADP-ribosylation is dynamic and takes place across many tissues undergoing renewal and repair, but the molecular mechanisms regulating this PTM remain mostly unknown. Here, we introduce the use of the planarian Schmidtea mediterranea as a tractable model to study PARylation in the complexity of the adult body that is under constant renewal and is capable of regenerating damaged tissues. We identified the evolutionary conservation of PARP signaling that is expressed in planarian stem cells and differentiated tissues. We also demonstrate that Smed-PARP-3 homolog is required for proper regeneration of tissues in the anterior region of the animal. Furthermore, our results demonstrate, Smed-PARP-3(RNAi) disrupts the timely location of injury-induced cell death near the anterior facing wounds and also affects the regeneration of the central nervous system. Our work reveals novel roles for PARylation in large-scale regeneration and provides a simplified platform to investigate PARP signaling in the complexity of the adult body.
Assuntos
Proteínas de Helminto/metabolismo , Planárias/fisiologia , Poli(ADP-Ribose) Polimerases/metabolismo , Regeneração/fisiologia , Animais , Morte Celular , Reparo do DNA/genética , Instabilidade Genômica , Proteínas de Helminto/antagonistas & inibidores , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Humanos , Neurogênese , Filogenia , Planárias/genética , Poli(ADP-Ribose) Polimerases/química , Poli(ADP-Ribose) Polimerases/classificação , Poli(ADP-Ribose) Polimerases/genética , Processamento de Proteína Pós-Traducional , Interferência de RNA , RNA de Cadeia Dupla/metabolismo , Transdução de SinaisRESUMO
Schistosoma mansoni venom allergen-like proteins (SmVALs) are part of a diverse protein superfamily partitioned into two groups (group 1 and group 2). Phylogenetic analyses of group 1 SmVALs revealed that members could be segregated into subclades (A-D); these subclades share similar gene expression patterns across the parasite lifecycle and immunological cross-reactivity. Furthermore, whole-mount in situ hybridization demonstrated that the phylogenetically, transcriptionally and immunologically-related SmVAL4, 10, 18 and 19 (subclade C) were all localized to the pre-acetabular glands of immature cercariae. Our results suggest that SmVAL group 1 phylogenetic relationships, stage-specific transcriptional profiles and tissue localization are predictive of immunological cross-reactivity.
Assuntos
Antígenos de Helmintos/imunologia , Proteínas de Helminto/imunologia , Filogenia , Schistosoma mansoni/química , Alérgenos/classificação , Alérgenos/genética , Alérgenos/imunologia , Animais , Antígenos de Helmintos/classificação , Antígenos de Helmintos/genética , Western Blotting , Reações Cruzadas , Vacinas contra Dengue/imunologia , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Perfilação da Expressão Gênica , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Soros Imunes/imunologia , Espectrometria de Massas , Família Multigênica , Schistosoma mansoni/classificação , Schistosoma mansoni/genética , Schistosoma mansoni/imunologia , Transcrição GênicaRESUMO
The characteristics of parasitic infections are often tied to host behavior. Although most studies have investigated definitive hosts, intermediate hosts can also play a role in shaping the distribution and accumulation of parasites. This is particularly relevant in larval stages, where intermediate host's behavior could potentially interfere in the molecules secreted by the parasite into the next host during infection. To investigate this hypothesis, we used a proteomic approach to analyze excretion/secretion products (ESP) from Fasciola hepatica newly excysted juveniles (NEJ) derived from two intermediate host species, Lymnaea viatrix and Pseudosuccinea columella. The two analyzed proteomes showed differences in identity, abundance, and functional classification of the proteins. This observation could be due to differences in the biological cycle of the parasite in the host, environmental aspects, and/or host-dependent factors. Categories such as protein modification machinery, protease inhibitors, signal transduction, and cysteine-rich proteins showed different abundance between samples. More specifically, differences in abundance of individual proteins such as peptidyl-prolyl cis-trans isomerase, thioredoxin, cathepsin B, cathepsin L, and Kunitz-type inhibitors were identified. Based on the differences identified between NEJ ESP samples, we can conclude that the intermediate host is a factor influencing the proteomic profile of ESP in F. hepatica.
Assuntos
Fasciola hepatica/metabolismo , Proteínas de Helminto/metabolismo , Lymnaea/parasitologia , Proteômica , Caramujos/parasitologia , Animais , Anidrases Carbônicas/classificação , Anidrases Carbônicas/metabolismo , Proteínas de Helminto/classificação , Larva/metabolismo , Peptídeo Hidrolases/classificação , Peptídeo Hidrolases/metabolismo , Peroxirredoxinas/classificação , Peroxirredoxinas/metabolismo , Inibidores de Proteases/classificação , Inibidores de Proteases/metabolismo , Receptores de Superfície Celular/classificação , Receptores de Superfície Celular/metabolismoRESUMO
We analyzed transcriptome profiles of Anisakis simplex (Nematoda: Anisakidae) 3rd (ASL3) and 4th larvae (ASL4) obtained by RNA-seq, to understand the molecular pathways linked to parasite survival and discover stage-enriched gene expressions. ASL3 were collected from chum salmon and ASL4 were obtained by in vitro culture. Whole transcriptome sequencing was conducted with Illumina sequencer, and de novo assembly was conducted. 47,179 and 41,934 genes were expressed in ASL3 and ASL4 transcriptomes. Of them, 17,633 were known and 29,546 were unmapped sequence for ASL3. 17,126 were known and 24,808 were unmapped sequence for ASL4. Polyubiquitins-related genes and collagen-related genes were the most abundantly expressed in ASL3 and ASL4. Mitochondrial enzyme-related genes were highly expressed both in ASL3 and ASL4. Among the transcripts, 675 were up-regulated in ASL3, while 1015 were up-regulated in ASL4. Several protease-related and protein biosynthesis-related genes were highly expressed in ASL3, all of which are thought to be crucial for invading host tissues. Collagen synthesis-related genes were highly expressed in ASL4, reflecting active biosynthesis of collagens during molting process. This information will extend our understanding of biology of the fish-borne zoonotic parasite A. simplex.
Assuntos
Anisaquíase/veterinária , Anisakis/genética , Doenças dos Peixes/parasitologia , Proteínas de Helminto/genética , Larva/genética , Oncorhynchus keta/parasitologia , Transcriptoma , Animais , Anisaquíase/parasitologia , Anisakis/classificação , Anisakis/crescimento & desenvolvimento , Colágeno/genética , Colágeno/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Larva/crescimento & desenvolvimento , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Anotação de Sequência Molecular , Filogenia , Poliubiquitina/genética , Poliubiquitina/metabolismoRESUMO
Despite the substantial amount of genomic and transcriptomic data available for a wide range of eukaryotic organisms, most genomes are still in a draft state and can have inaccurate gene predictions. To gain a sound understanding of the biology of an organism, it is crucial that inferred protein sequences are accurately identified and annotated. However, this can be challenging to achieve, particularly for organisms such as parasitic worms (helminths), as most gene prediction approaches do not account for substantial phylogenetic divergence from model organisms, such as Caenorhabditis elegans and Drosophila melanogaster, whose genomes are well-curated. In this paper, we describe a bioinformatic strategy for the curation of gene families and subsequent annotation of encoded proteins. This strategy relies on pairwise gene curation between at least two closely related species using genomic and transcriptomic data sets, and is built on recent work on kinase complements of parasitic worms. Here, we discuss salient technical aspects of this strategy and its implications for the curation of protein families more generally.
Assuntos
Genoma Helmíntico , Haemonchus/genética , Proteínas de Helminto/genética , Proteínas Quinases/genética , Schistosoma/genética , Trichinella/genética , Trichuris/genética , Animais , Caenorhabditis elegans/classificação , Caenorhabditis elegans/enzimologia , Caenorhabditis elegans/genética , Biologia Computacional/métodos , Curadoria de Dados/métodos , Bases de Dados Genéticas , Feminino , Ontologia Genética , Haemonchus/classificação , Haemonchus/enzimologia , Proteínas de Helminto/classificação , Proteínas de Helminto/metabolismo , Anotação de Sequência Molecular/métodos , Filogenia , Proteínas Quinases/classificação , Proteínas Quinases/metabolismo , Schistosoma/classificação , Schistosoma/enzimologia , Transcriptoma , Trichinella/classificação , Trichinella/enzimologia , Trichuris/classificação , Trichuris/enzimologiaRESUMO
Schistosomes infect more than 200 million people. These parasitic flatworms rely on a syncytial outer coat called the tegument to survive within the vasculature of their host. Although the tegument is pivotal for their survival, little is known about maintenance of this tissue during the decades schistosomes survive in the bloodstream. Here, we demonstrate that the tegument relies on stem cells (neoblasts) to specify fusogenic progenitors that replace tegumental cells lost to turnover. Molecular characterization of neoblasts and tegumental progenitors led to the discovery of two flatworm-specific zinc finger proteins that are essential for tegumental cell specification. These proteins are homologous to a protein essential for neoblast-driven epidermal maintenance in free-living flatworms. Therefore, we speculate that related parasites (i.e., tapeworms and flukes) employ similar strategies to control tegumental maintenance. Since parasitic flatworms infect every vertebrate species, understanding neoblast-driven tegumental maintenance could identify broad-spectrum therapeutics to fight diseases caused by these parasites.
Assuntos
Regulação da Expressão Gênica , Platelmintos/genética , Schistosoma mansoni/genética , Células-Tronco/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Células Epidérmicas/citologia , Células Epidérmicas/metabolismo , Células Epidérmicas/parasitologia , Epiderme/metabolismo , Epiderme/parasitologia , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Filogenia , Platelmintos/citologia , Platelmintos/fisiologia , Interferência de RNA , Esquistossomose mansoni/parasitologia , Homologia de Sequência de AminoácidosRESUMO
Strongyloides spp., gastrointestinal nematode parasites of humans and other animals, have genetically identical parasitic and free-living adult life cycle stages. This is an almost unique feature amongst nematodes and comparison of these two stages can provide insights into the genetic basis and evolution of Strongyloides nematode parasitism. Here, we present RNAseq data for S. venezuelensis, a parasite of rodents, and identify genes that are differentially expressed in parasitic and free-living life cycle stages. Comparison of these data with analogous RNAseq data for three other Strongyloides spp., has identified key protein-coding gene families with a putative role in parasitism including WAGO-like Argonautes (at the genus level) and speckle-type POZ-like coding genes (S. venezuelensis-S. papillosus phylogenetic subclade level). Diverse gene families are uniquely upregulated in the parasitic stage of all four Strongyloides species, including a distinct upregulation of genes encoding cytochrome P450 in S. venezuelensis, suggesting some diversification of the molecular tools used in the parasitic life cycle stage among individual species. Together, our results identify key gene families with a putative role in Strongyloides parasitism or features of the parasitic life cycle stage, and deepen our understanding of parasitism evolution among Strongyloides species.
Assuntos
Filogenia , Strongyloides/genética , Estrongiloidíase/genética , Transcriptoma/genética , Animais , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Humanos , Larva/genética , Larva/patogenicidade , Estágios do Ciclo de Vida/genética , Ratos Wistar , Roedores/parasitologia , Strongyloides/patogenicidade , Estrongiloidíase/parasitologia , Simbiose/genéticaRESUMO
Mechanisms underlying anteroposterior body axis differences during adult tissue maintenance and regeneration are poorly understood. Here, we identify that post-translational modifications through the SUMO (Small Ubiquitin-like Modifier) machinery are evolutionarily conserved in the Lophotrocozoan Schmidtea mediterranea. Disruption of SUMOylation in adult animals by RNA-interference of the only SUMO E2 conjugating enzyme Ubc9 leads to a systemic increase in DNA damage and a remarkable regional defect characterized by increased cell death and loss of the posterior half of the body. We identified that Ubc9 is mainly expressed in planarian stem cells (neoblasts) but it is also transcribed in differentiated cells including neurons. Regeneration in Ubc9(RNAi) animals is impaired and associated with low neoblast proliferation. We present evidence indicating that Ubc9-induced regional cell death is preceded by alterations in transcription and spatial expression of repressors and activators of the Hedgehog signaling pathway. Our results demonstrate that SUMOylation acts as a regional-specific cue to regulate cell fate during tissue renewal and regeneration.
Assuntos
Proliferação de Células , Proteínas Hedgehog/metabolismo , Proteínas de Helminto/metabolismo , Planárias/metabolismo , Transdução de Sinais , Células-Tronco/metabolismo , Sequência de Aminoácidos , Animais , Morte Celular , Proteínas Hedgehog/genética , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Filogenia , Planárias/citologia , Planárias/genética , Interferência de RNA , Homologia de Sequência de Aminoácidos , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/classificação , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/genética , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Células-Tronco/citologia , Sumoilação , Enzimas de Conjugação de Ubiquitina/classificação , Enzimas de Conjugação de Ubiquitina/genética , Enzimas de Conjugação de Ubiquitina/metabolismoRESUMO
The hnRNPs play important roles in physiological processes in eukaryotic organisms by regulation of pre-mRNA after transcription, including pre-mRNA splicing, mRNA stability, DNA replication and repair and telomere maintenance and so on. However, it remains unclear about the specific functions of these genes. In this study, the full-length cDNA sequence of hnRNPA2/B1-like was first cloned from Dugesia japonica, and its roles were investigated by WISH and RNAi. The results showed that: (1) DjhnRNPA2/B1-like was highly conserved during animal evolution; (2) DjhnRNPA2/B1-like mRNA was mainly distributed each side of the body in intact worms and regenerative blastemas, and its expression levels were up-regulated on days 0 and 5 after amputation; (3) the intact and regenerating worms gradually lysed or lost regeneration capacity after DjhnRNPA2/B1-like RNAi; and (4) DjhnRNPA2/B1-like expression is induced by temperature and heavy metal ion stress. The data suggests that DjhnRNPA2/B1-like is a multiple functional gene, it plays important roles in regeneration and homeostatic maintenance and it is also involved in stress responses in planarians. Our work provides basic data for the study of regenerative mechanism and stress responses in freshwater planarians.
Assuntos
Proteínas de Helminto/fisiologia , Ribonucleoproteínas Nucleares Heterogêneas/fisiologia , Homeostase/genética , Planárias/genética , Planárias/fisiologia , Regeneração/genética , Animais , DNA Complementar/genética , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Ribonucleoproteínas Nucleares Heterogêneas/classificação , Ribonucleoproteínas Nucleares Heterogêneas/genética , Hibridização In Situ , Metais Pesados/toxicidade , Interferência de RNA/fisiologia , Precursores de RNA/metabolismo , RNA Mensageiro/metabolismo , Homologia de Sequência de Aminoácidos , Estresse Fisiológico/genética , TemperaturaRESUMO
Fasciola gigantica is regarded as the major liver fluke causing fasciolosis in livestock in tropical countries. Despite the significant economic and public health impacts of F. gigantica there are few studies on the pathogenesis of this parasite and our understanding is further limited by the lack of genome and transcriptome information. In this study, de novo Illumina RNA sequencing (RNA-seq) was performed to obtain a comprehensive transcriptome profile of the juvenile (42days post infection) and adult stages of F. gigantica. A total of 49,720 unigenes were produced from juvenile and adult stages of F. gigantica, with an average length of 1286 nucleotides (nt) and N50 of 2076nt. A total of 27,862 (56.03%) unigenes were annotated by BLAST similarity searches against the NCBI non-redundant protein database. Because F. gigantica needs to feed and/or digest host tissues, some proteases (including cysteine proteases and aspartic proteases), which play a role in the degradation of host tissues (protein), have been paid more attention in the present study. A total of 6511 distinct genes were found differentially expressed between juveniles and adults, of which 3993 genes were up-regulated and 2518 genes were down-regulated in adults versus juveniles, respectively. Moreover, stage-specific differentially expressed genes were identified in juvenile (17,009) and adult (6517) F. gigantica. The significantly divergent pathways of differentially expressed genes included cAMP signaling pathway (226; 4.12%), proteoglycans in cancer (256; 4.67%) and focal adhesion (199; 3.63%). The transcription pattern also revealed two egg-laying-associated pathways: cGMP-PKG signaling pathway and TGF-ß signaling pathway. This study provides the first comparative transcriptomic data concerning juvenile and adult stages of F. gigantica that will be of great value for future research efforts into understanding parasite pathogenesis and developing vaccines against this important parasite.
Assuntos
Fasciola/genética , Fasciolíase/veterinária , Regulação da Expressão Gênica no Desenvolvimento , Genes de Helmintos , Proteínas de Helminto/genética , Redes e Vias Metabólicas/genética , Transcriptoma , Animais , Ácido Aspártico Proteases/classificação , Ácido Aspártico Proteases/genética , Búfalos , Cisteína Proteases/classificação , Cisteína Proteases/genética , Bases de Dados Genéticas , Fasciola/isolamento & purificação , Fasciola/metabolismo , Fasciolíase/parasitologia , Perfilação da Expressão Gênica , Ontologia Genética , Proteínas de Helminto/classificação , Sequenciamento de Nucleotídeos em Larga Escala , Anotação de Sequência Molecular , Transdução de SinaisRESUMO
The root-knot nematode, Meloidogyne incognita causes significant damage to various economically important crops. Infection is associated with secretion of effector proteins into host cytoplasm and interference with host innate immunity. To combat this infection, the identification and functional annotations of Excretory/Secretory (ES) proteins serve as a key to produce durable control measures. The identification of ES proteins through experimental methods are expensive and time consuming while bioinformatics approaches are cost-effective by prioritizing the experimental analysis of potential drug targets for parasitic diseases. In this study, we predicted and functionally annotated the 1889 ES proteins in M. incognita genome using integration of several bioinformatics tools. Of these 1889 ES proteins, 473 (25%) had orthologues in free living nematode Caenorhabditis elegans, 825(67.8%) in parasitic nematodes whereas 561 (29.7%) appeared to be novel and M. incognita specific molecules. Of the C. elegans homologues, 17 ES proteins had "loss of function phenotype" by RNA interference and could represent potential drug targets for parasite intervention and control. We could functionally annotate 429 (22.7%) ES proteins using Gene Ontology (GO) terms, 672 (35.5%) proteins to protein domains and established pathway associations for 223 (11.8%) sequences using Kyoto Encyclopaedia of Genes and Genomes (KEGG). The 162 (8.5%) ES proteins were also mapped to several important plant cell-wall degrading CAZyme families including chitinase, cellulase, xylanase, pectate lyase and endo-ß-1,4-xylanase. Our comprehensive analysis of M. incognita secretome provides functional information for further experimental study.
Assuntos
Genoma Helmíntico/genética , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Proteoma/classificação , Proteoma/genética , Animais , Biologia Computacional , Feminino , Ontologia Genética , Masculino , Domínios Proteicos , Tylenchoidea/genéticaRESUMO
BACKGROUND: Proteins of the cysteine-rich secretory proteins, antigen 5 and pathogenesis-related 1 (CAP) superfamily are recognized or proposed to play roles in parasite development and reproduction, and in modulating host immune attack and infection processes. However, little is known about these proteins for most parasites. RESULTS: In the present study, we explored CAP proteins of Toxocara canis, a socioeconomically important zoonotic roundworm. To do this, we mined and curated transcriptomic and genomic data, predicted and curated full-length protein sequences (n = 28), conducted analyses of these data and studied the transcription of respective genes in different developmental stages of T. canis. In addition, based on information available for Caenorhabditis elegans, we inferred that selected genes (including lon-1, vap-1, vap-2, scl-1, scl-8 and scl-11 orthologs) of T. canis and their interaction partners likely play central roles in this parasite's development and/or reproduction via TGF-beta and/or insulin-like signaling pathways, or via host interactions. CONCLUSION: In conclusion, this study could provide a foundation to guide future studies of CAP proteins of T. canis and related parasites, and might assist in finding new interventions against diseases caused by these parasites.