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1.
Gene ; 806: 145929, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34461150

RESUMO

The body color of Neocaridina denticulate sinensis is a compelling phenotypic trait, in which a cascade of carotenoid metabolic processes plays an important role. The study was conducted to compare the transcriptome of cephalothoraxes among three pigmentation phenotypes (red, blue, and chocolate) of N. denticulate sinensis. The purpose of this study was to explore the candidate genes associated with different colors of N. denticulate sinensis. Nine cDNA libraries in three groups were constructed from the cephalothoraxes of shrimps. After assembly, 75022 unigenes were obtained in total with an average length of 1026 bp and N50 length of 1876 bp. There were 45977, 25284, 23605, 21913 unigenes annotated in the Nr, Swissprot, KOG, and KEGG databases, respectively. Differential expression analysis revealed that there were 829, 554, and 3194 differentially expressed genes (DEGs) in RD vs BL, RD vs CH, and BL vs CH, respectively. These DEGs may play roles in the absorption, transport, and metabolism of carotenoids. We also emphasized that electron transfer across the inner mitochondrial membrane (IMM) was a key process in pigment metabolism. In addition, a total of 6328 simple sequence repeats (SSRs) were also detected in N. denticulate sinensis. The results laid a solid foundation for further research on the molecular mechanism of integument pigmentation in the crustacean and contributed to developing more attractive aquatic animals.


Assuntos
Proteínas de Artrópodes/genética , Carotenoides/metabolismo , Decápodes/genética , Pigmentação/genética , Transcriptoma , Animais , Organismos Aquáticos , Proteínas de Artrópodes/classificação , Proteínas de Artrópodes/metabolismo , Transporte Biológico , Cor , Bases de Dados Genéticas , Decápodes/anatomia & histologia , Decápodes/metabolismo , Água Doce , Regulação da Expressão Gênica , Biblioteca Gênica , Ontologia Genética , Repetições de Microssatélites , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Anotação de Sequência Molecular , Característica Quantitativa Herdável
2.
Int J Mol Sci ; 22(16)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34445418

RESUMO

Central pattern generators produce rhythmic behaviors independently of sensory input; however, their outputs can be modulated by neuropeptides, thereby allowing for functional flexibility. We investigated the effects of C-type allatostatins (AST-C) on the cardiac ganglion (CG), which is the central pattern generator that controls the heart of the American lobster, Homarus americanus, to identify the biological mechanism underlying the significant variability in individual responses to AST-C. We proposed that the presence of multiple receptors, and thus differential receptor distribution, was at least partly responsible for this observed variability. Using transcriptome mining and PCR-based cloning, we identified four AST-C receptors (ASTCRs) in the CG; we then characterized their cellular localization, binding potential, and functional activation. Only two of the four receptors, ASTCR1 and ASTCR2, were fully functional GPCRs that targeted to the cell surface and were activated by AST-C peptides in our insect cell expression system. All four, however, were amplified from CG cDNAs. Following the confirmation of ASTCR expression, we used physiological and bioinformatic techniques to correlate receptor expression with cardiac responses to AST-C across individuals. Expression of ASTCR1 in the CG showed a negative correlation with increasing contraction amplitude in response to AST-C perfusion through the lobster heart, suggesting that the differential expression of ASTCRs within the CG is partly responsible for the specific physiological response to AST-C exhibited by a given individual lobster.


Assuntos
Perfilação da Expressão Gênica/métodos , Nephropidae/genética , Neuropeptídeos/farmacologia , Receptores de Neuropeptídeos/genética , Receptores de Neuropeptídeos/metabolismo , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Sistema Cardiovascular/metabolismo , Membrana Celular/metabolismo , Clonagem Molecular , Mineração de Dados , Bases de Dados Genéticas , Regulação da Expressão Gênica/efeitos dos fármacos , Miocárdio/metabolismo , Nephropidae/efeitos dos fármacos , Nephropidae/metabolismo , Análise de Sequência de RNA , Células Sf9 , Distribuição Tecidual
3.
Int J Mol Sci ; 22(12)2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34208769

RESUMO

Early changes in hemocyte proteins in freshwater crayfish Pacifastacus leniusculus, in response to an injection with the fungal pattern recognition protein ß-1,3-glucan (laminarin) were investigated, as well as changes after saline (vehicle) injection and in naïve animals. Injection of saline resulted in rapid recruitment of granular hemocytes from surrounding tissues, whereas laminarin injection on the other hand induced an initial dramatic drop of hemocytes. At six hours after injection, the hemocyte populations therefore were of different composition. The results show that mature granular hemocytes increase in number after saline injection as indicated by the high abundance of proteins present in granular cell vesicles, such as a vitelline membrane outer layer protein 1 homolog, mannose-binding lectin, masquerade, crustin 1 and serine protease homolog 1. After injection with the ß-1,3-glucan, only three proteins were enhanced in expression, in comparison with saline-injected animals and uninjected controls. All of them may be associated with immune responses, such as a new and previously undescribed Kazal proteinase inhibitor. One interesting observation was that the clotting protein was increased dramatically in most of the animals injected with laminarin. The number of significantly affected proteins was very few after a laminarin injection when compared to uninjected and saline-injected crayfish. This finding may demonstrate some problematic issues with gene and protein expression studies from other crustaceans receiving injections with pathogens or pattern recognition proteins. If no uninjected controls are included and no information about hemocyte count (total or differential) is given, expressions data for proteins or mRNAs are very difficult to properly interpret.


Assuntos
Astacoidea/efeitos dos fármacos , Astacoidea/metabolismo , Hemócitos/metabolismo , beta-Glucanas/administração & dosagem , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Astacoidea/genética , Biomarcadores , Expressão Gênica , Hemócitos/citologia , Proteoma , Proteômica/métodos , RNA Mensageiro/genética
4.
Commun Biol ; 4(1): 668, 2021 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-34083730

RESUMO

Diet is a powerful evolutionary force for species adaptation and diversification. Acari is one of the most abundant clades of Arachnida, exhibiting diverse dietary types, while the underlying genetic adaptive mechanisms are not fully understood. Based on comparative analyses of 15 Acari genomes, we found genetic bases for three specialized diets. Herbivores experienced stronger selection pressure than other groups; the olfactory genes and gene families involving metabolizing toxins showed strong adaptive signals. Genes and gene families related to anticoagulation, detoxification, and haemoglobin digestion were found to be under strong selection pressure or significantly expanded in the blood-feeding species. Lipid metabolism genes have a faster evolutionary rate and been subjected to greater selection pressures in fat-feeding species; one positively selected site in the fatty-acid amide hydrolases 2 gene was identified. Our research provides a new perspective for the evolution of Acari and offers potential target loci for novel pesticide development.


Assuntos
Adaptação Fisiológica/genética , Dieta , Genoma/genética , Ácaros/genética , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Evolução Molecular , Variação Genética , Herbivoria/genética , Humanos , Ácaros/classificação , Ácaros/metabolismo , Filogenia , Seleção Genética , Homologia de Sequência de Aminoácidos , Especificidade da Espécie
5.
Int J Biol Macromol ; 184: 821-830, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34171258

RESUMO

dUTPases are key enzymes in all life kingdoms. A staphylococcal repressor protein (Stl) inhibited dUTPases from multiple species to various extents. Understanding the molecular basis underlying the inhibition differences is crucial to develop effective proteinaceous inhibitors of dUTPases. Herein, we report the complex structure of Stl N-terminal domain (StlN-ter) and Litopenaeus vannamei dUTPase domain (lvDUT65-210). Stl inhibited lvDUT65-210 through its N-terminal domain. The lvDUT65-210-StlN-ter complex structure revealed a heterohexamer encompassing three StlN-ter monomers bound to one lvDUT65-210 trimer, generating two types of Stl-dUTPase interfaces. Interface I is formed by Stl interaction with the lvDUT65-210 active-site region that is contributed by motifs I-IV from its two subunits; interface II results from Stl binding to the C-terminal motif V of the third lvDUT65-210 subunit. Structural comparison revealed both conserved features and obvious differences in Stl-dUTPase interaction patterns, giving clues about the inhibition differences of Stl on dUTPases. Noticeably, interface II is only observed in lvDUT65-210-StlN-ter. The Stl-interacting residues of lvDUT65-210 are conserved in other eukaryotic dUTPases, particularly human dUTPase. Altogether, our study presents the first structural model of Stl interaction with eukaryotic dUTPase, contributing to a more complete view of Stl inhibition and facilitating the development of proteinaceous inhibitor for eukaryotic dUTPases.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Penaeidae/enzimologia , Pirofosfatases/química , Pirofosfatases/metabolismo , Staphylococcus aureus/metabolismo , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Modelos Moleculares , Penaeidae/química , Ligação Proteica , Conformação Proteica , Domínios Proteicos , Multimerização Proteica , Staphylococcus aureus/química
6.
Viruses ; 13(4)2021 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-33921873

RESUMO

Coronavirus-like organisms have been previously identified in Arthropod ectoparasites (such as ticks and unfed cat flea). Yet, the question regarding the possible role of these arthropods as SARS-CoV-2 passive/biological transmission vectors is still poorly explored. In this study, we performed in silico structural and binding energy calculations to assess the risks associated with possible ectoparasite transmission. We found sufficient similarity between ectoparasite ACE and human ACE2 protein sequences to build good quality 3D-models of the SARS-CoV-2 Spike:ACE complex to assess the impacts of ectoparasite mutations on complex stability. For several species (e.g., water flea, deer tick, body louse), our analyses showed no significant destabilisation of the SARS-CoV-2 Spike:ACE complex, suggesting these species would bind the viral Spike protein. Our structural analyses also provide structural rationale for interactions between the viral Spike and the ectoparasite ACE proteins. Although we do not have experimental evidence of infection in these ectoparasites, the predicted stability of the complex suggests this is possible, raising concerns of a possible role in passive transmission of the virus to their human hosts.


Assuntos
Proteínas de Artrópodes/metabolismo , Artrópodes/metabolismo , Peptidil Dipeptidase A/metabolismo , SARS-CoV-2/metabolismo , Enzima de Conversão de Angiotensina 2/química , Enzima de Conversão de Angiotensina 2/genética , Enzima de Conversão de Angiotensina 2/metabolismo , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Artrópodes/química , Artrópodes/classificação , Artrópodes/genética , Sítios de Ligação , COVID-19/transmissão , Ectoparasitoses/parasitologia , Humanos , Modelos Moleculares , Mutação , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/genética , Filogenia , Ligação Proteica , SARS-CoV-2/química , SARS-CoV-2/genética , Homologia de Sequência , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/metabolismo
7.
Int J Mol Sci ; 22(7)2021 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-33806079

RESUMO

We focus on the stalked goose barnacle L. anatifera adhesive system, an opportunistic less selective species for the substrate, found attached to a variety of floating objects at seas. Adhesion is an adaptative character in barnacles, ensuring adequate positioning in the habitat for feeding and reproduction. The protein composition of the cement multicomplex and adhesive gland was quantitatively studied using shotgun proteomic analysis. Overall, 11,795 peptide sequences were identified in the gland and 2206 in the cement, clustered in 1689 and 217 proteinGroups, respectively. Cement specific adhesive proteins (CPs), proteases, protease inhibitors, cuticular and structural proteins, chemical cues, and many unannotated proteins were found, among others. In the cement, CPs were the most abundant (80.5%), being the bulk proteins CP100k and -52k the most expressed of all, and CP43k-like the most expressed interfacial protein. Unannotated proteins comprised 4.7% of the cement proteome, ranking several of them among the most highly expressed. Eight of these proteins showed similar physicochemical properties and amino acid composition to known CPs and classified through Principal Components Analysis (PCA) as new CPs. The importance of PCA on the identification of unannotated non-conserved adhesive proteins, whose selective pressure is on their relative amino acid abundance, was demonstrated.


Assuntos
Adesivos , Peptídeos/metabolismo , Proteogenômica , Proteoma , Thoracica/metabolismo , Animais , Proteínas de Artrópodes/metabolismo , Análise por Conglomerados , Ecossistema , Peso Molecular , Análise de Componente Principal , Proteômica/métodos
8.
J Proteome Res ; 20(5): 2923-2934, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33851848

RESUMO

Hypoxia is one of the major stresses in aquaculture animals. Recently, we reported that hypoxia disrupts the endocrine system and inhibits testicular function of oriental river prawns (Macrobrachium nipponense), but the molecular mechanism of testes responded to hypoxia remains largely unknown. In the present study, we aimed to integrate whole phosphoproteomic profiles of hypoxia-treated testes of the oriental river prawn (Macrobrachium nipponense). We successfully isolated sperm cells and evaluated the mitochondrial morphology and function using laser confocal microscopy, flow cytometry, and biochemical analyses. Quantitative proteomics identified 117 differentially abundant phosphorylated proteins, and these proteins are mainly involved in the pathways related to cellular processes, including autophagy, apoptosis, and the FoxO signaling pathway. Protein-protein interaction analysis clustered these phosphoproteins into three groups, many of which have been suggested to impact carbohydrate metabolism, autophagy, and signal regulation in testes. Western blotting confirmed that phosphorylated proteins including AMPK, ULK1, and TP53 (of the AMPK pathway) may contribute to testicular dysfunction caused by hypoxia. Further, we investigated the potential roles of AMP-activated protein kinase (AMPK)'s in testes mitochondrial autophagy and apoptosis in M. nipponense as induced by hypoxia. Simultaneous knockdown of AMPKα in sperm cells led to a decrease in FOXO3a phosphorylation at Ser413, upregulation of caspase-3 and caspase-9 activities, and an increased apoptosis rate. These results improve our understanding of hypoxia-induced energy metabolism disorders in the testes of M. nipponense.


Assuntos
Palaemonidae , Proteínas Quinases Ativadas por AMP/genética , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Regulação da Expressão Gênica , Hipóxia , Masculino , Palaemonidae/genética , Palaemonidae/metabolismo , Transdução de Sinais , Testículo/metabolismo
9.
Int J Biol Macromol ; 183: 707-717, 2021 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-33930448

RESUMO

Akirin is a highly conserved nuclear factor among different species. It is closely related to skeletal muscle development, innate immune response, and tumorigenesis in a variety of animals. In invertebrates, Akirin is mainly involved in gene transcription and NF-κB dependent natural immune response. In the present study, a nuclear factor Akirin was identified from Procambarus clarkii. The Akirin protein of crayfish consists of 204 amino acids and is conserved among its family members, especially the nuclear localization signal peptide motif (KRRR). PcAkirin was highly expressed in stomach, intestines, and hepatopancreas. After A. hydrophila challenge, the transcription level of Akirin significantly increased in hemocyte and hepatopancreas. In addition, the recombinant Akirin protein was produced successfully and helpful to resist WSSV infection by increasing the expression level of some immune related genes. On the contrary, after interfering with Akirin gene by dsRNA, the crayfish increased the sensitivity to A. hydrophila and WSSV infections. The results are more obvious in the accumulated mortality of P. clarkii infected with A. hydrophila and WSSV. All these results suggested that Akirin played a significant role in innate immune responses and protected it from WSSV and bacterial infection in crayfish.


Assuntos
Astacoidea/virologia , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Citotóxicas Formadoras de Poros/genética , Vírus da Síndrome da Mancha Branca 1/patogenicidade , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Astacoidea/imunologia , Clonagem Molecular , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Imunidade Inata , Distribuição Tecidual , Vírus da Síndrome da Mancha Branca 1/imunologia
10.
J Fish Dis ; 44(8): 1131-1145, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33835515

RESUMO

Whiteleg shrimp is a widely cultured crustacean, but frequent disease outbreaks have decreased production and caused significant losses. Toll-like receptors (TLRs) comprise a large innate immune family that is involved in the innate immune response. However, understanding of their regulatory mechanism is limited. In this study, PacBio sequencing and Illumina sequencing were applied to the gill and hepatopancreas tissues of whiteleg shrimp and an integrated transcript gene set was established. The upregulation of Toll1, Toll2 and Toll3 transcripts in the hepatopancreas tissue of whiteleg shrimp after iridescent virus infection implies that these proteins are involved in the immune response to the virus; simultaneously, the TRAF6 and relish transcripts in the Toll pathway were also upregulated, implying that the Toll pathway was activated. We predicted the three-dimensional structure of the five Toll proteins in whiteleg shrimp and humans and constructed a phylogenetic tree of the Toll protein family. In addition, there was a large discrepancy of Toll1 between invertebrates and vertebrates, presumably because of the loss of Toll1 protein sequence during the evolution process from invertebrates to vertebrates. Our research will improve the cognition of Toll protein family in invertebrates in terms of evolution, structure and function and provide theoretical guidance for researchers in this field.


Assuntos
Proteínas de Artrópodes/genética , Evolução Molecular , Iridoviridae/fisiologia , Penaeidae/genética , Receptores Toll-Like/genética , Animais , Proteínas de Artrópodes/metabolismo , Penaeidae/virologia , Análise de Sequência de DNA , Análise de Sequência de Proteína , Receptores Toll-Like/metabolismo , Transcrição Genética
11.
Gene ; 788: 145583, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33753150

RESUMO

Macrobrachium nipponense has the characteristics of fast ovarian development cycle, which leads to the coexistence of multiple generations, the reduction of commodity specifications and the low economic benefit. Therefore, the study on the mechanism of ovarian development is of great significance to the development of industry. Cyclin A (CycA)is a key gene regulating ovarian development in vertebrates, but little information was available for its function in crustaceans. In this study, the full-length cDNA of Mn-CycA was obtained from the ovary. The full-length cDNA (2033 bp) with an open reading frame of 1368 bp, encoded a 456-amino acid protein. qRT-PCR revealed tissue-specific expression pattern of Mn-CycA, with abundant expression in the ovary. Results in different developmental stages of ovary indicated that Mn-CycA expression is positively correlated with ovarian maturation. qRT-PCR In different developmental stages, the expression of Mn-CycA mRNA gradually increased during the embryonic stage and decreased significantly on the first day of the hatching stage. At the 25th day of the metamorphosis stage, the expression level of Mn-CycAmRNA in female shrimp was 3.5 times higher than that in male shrimp, which may be related to the proliferation of oogonia and the formation of oocytes. In situ hybridization (ISH) of ovary showed Mn-CycA was examined in all stages and was mainly located in oogonia and oocytes. Compared with the control group, the obvious change of gonad somatic index (GSI) proved that injection of Mn-CycA dsRNA could delay the ovarian development cycle, which provided strong evidence for the involvement of Mn-CycA in ovarian maturation and oogenesis, and expanded a new perspective for studying the fast ovarian development cycle in M. nipponense.


Assuntos
Ciclina A/genética , Ciclina A/metabolismo , Perfilação da Expressão Gênica/métodos , Palaemonidae/crescimento & desenvolvimento , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Sequência de Bases , Clonagem Molecular , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogônios/crescimento & desenvolvimento , Oogônios/metabolismo , Fases de Leitura Aberta , Especificidade de Órgãos , Palaemonidae/genética , Palaemonidae/metabolismo , Filogenia
12.
Parasit Vectors ; 14(1): 144, 2021 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-33676573

RESUMO

BACKGROUND: Louping ill virus (LIV) and tick-borne encephalitis virus (TBEV) are tick-borne flaviviruses that are both transmitted by the major European tick, Ixodes ricinus. Despite the importance of I. ricinus as an arthropod vector, its capacity to acquire and subsequently transmit viruses, known as vector competence, is poorly understood. At the molecular scale, vector competence is governed in part by binary interactions established between viral and cellular proteins within infected tick cells. METHODS: To investigate virus-vector protein-protein interactions (PPIs), the entire set of open reading frames for LIV and TBEV was screened against an I. ricinus cDNA library established from three embryonic tick cell lines using yeast two-hybrid methodology (Y2H). PPIs revealed for each viral bait were retested in yeast by applying a gap repair (GR) strategy, and notably against the cognate protein of both viruses, to determine whether the PPIs were specific for a single virus or common to both. The interacting tick proteins were identified by automatic BLASTX, and in silico analyses were performed to expose the biological processes targeted by LIV and TBEV. RESULTS: For each virus, we identified 24 different PPIs involving six viral proteins and 22 unique tick proteins, with all PPIs being common to both viruses. According to our data, several viral proteins (pM, M, NS2A, NS4A, 2K and NS5) target multiple tick protein modules implicated in critical biological pathways. Of note, the NS5 and pM viral proteins establish PPI with several tumor necrosis factor (TNF) receptor-associated factor (TRAF) proteins, which are essential adaptor proteins at the nexus of multiple signal transduction pathways. CONCLUSION: We provide the first description of the TBEV/LIV-I. ricinus PPI network, and indeed of any PPI network involving a tick-borne virus and its tick vector. While further investigation will be needed to elucidate the role of each tick protein in the replication cycle of tick-borne flaviviruses, our study provides a foundation for understanding the vector competence of I. ricinus at the molecular level. Indeed, certain PPIs may represent molecular determinants of vector competence of I. ricinus for TBEV and LIV, and potentially for other tick-borne flaviviruses.


Assuntos
Proteínas de Artrópodes/metabolismo , Vírus da Encefalite Transmitidos por Carrapatos/genética , Vírus da Encefalite Transmitidos por Carrapatos/fisiologia , Interações entre Hospedeiro e Microrganismos , Ixodes/genética , Ixodes/virologia , Proteínas Virais/metabolismo , Animais , Proteínas de Artrópodes/genética , Feminino , Biblioteca Gênica , Fases de Leitura Aberta , Domínios e Motivos de Interação entre Proteínas , Proteínas Virais/genética
13.
Biochim Biophys Acta Proteins Proteom ; 1869(6): 140642, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33647452

RESUMO

Anhydrobiotic organisms accumulate late embryogenesis abundant (LEA) proteins, a family of intrinsically disordered proteins (IDPs) reported to improve cellular tolerance to water stress. Here we show that AfrLEA6, a Group 6 LEA protein only recently discovered in animals, protects lactate dehydrogenase (LDH), citrate synthase (CS) and phosphofructokinase (PFK) against damage during desiccation. In some cases, protection is enhanced by trehalose, a naturally-occurring protective solute. An open question is whether gain of secondary structure by LEA proteins during drying is a prerequisite for this stabilizing function. We used incremental drying (equilibration to a series of relative humidities, RH) to test the ability of AfrLEA2, a Group 3 LEA protein, to protect desiccation-sensitive PFK. AfrLEA2 was chosen due to its exceptional ability to protect PFK. In parallel, circular dichroism (CD) spectra were obtained for AfrLEA2 across the identical range of relative water contents. Protection of PFK by AfrLEA2, above that observed with trehalose and BSA, coincides with simultaneous gain of α-helix in AfrLEA2. At 100% RH, the CD spectrum for AfrLEA2 is typical of random coil, while at decreasing RH, the spectrum shows higher ellipticity at 191 nm and minima at 208 and 220 nm, diagnostic of α-helix. This study provides experimental evidence linking the gain of α-helix with stabilization of a target protein across a graded series of hydration states. Mechanistically, it is intriguing that certain other functions of these IDPs, like preventing aggregation of target proteins, can occur in fully hydrated cells and apparently do not require gain of α-helix.


Assuntos
Artemia/metabolismo , Proteínas Intrinsicamente Desordenadas/química , Proteínas Intrinsicamente Desordenadas/metabolismo , Fosfofrutoquinases/metabolismo , Animais , Artemia/química , Proteínas de Artrópodes/química , Proteínas de Artrópodes/metabolismo , Dicroísmo Circular , Dessecação , Fosfofrutoquinases/química , Conformação Proteica em alfa-Hélice , Dobramento de Proteína , Estabilidade Proteica
14.
Toxins (Basel) ; 13(2)2021 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-33673184

RESUMO

Latroeggtoxin-VI (LETX-VI) is a peptide neurotoxin newly found from the eggs of spider L. tredecimguttatus. To explore the mechanism of action of the LETX-VI on nerve cells, the effects of LETX-VI on PC12 cells, a commonly used neuron model, were analyzed using a pull-down assay-guided strategy. LETX-VI was shown to interact with 164 PC12 cell proteins that have diverse molecular functions such as binding, catalysis, regulation, structural activity, etc., thereby extensively affecting the biological processes in the PC12 cells, particularly protein metabolism, response to stimulus, substance transport, and nucleic acid metabolism, with 56.71%, 42.07%, 29.88% and 28.66% of the identified proteins being involved in these biological processes, respectively. By interacting with the relevant proteins, LETX-VI enhanced the synthesis of dopamine; positively regulated cell division and proliferation; and negatively regulated cell cycle arrest, cell death, and apoptotic processes, and therefore has limited cytotoxicity against the PC12 cells, which were further experimentally confirmed. In general, the effects of LETX-VI on PC12 cells are more regulatory than cytotoxic. These findings have deepened our understanding of the action mechanism of LETX-VI on nerve cells and provided valuable clues for further related researches including those on Parkinson's disease.


Assuntos
Proteínas de Artrópodes/toxicidade , Neurônios Dopaminérgicos/efeitos dos fármacos , Mapeamento de Interação de Proteínas , Proteoma , Proteômica , Animais , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Ciclo Celular/efeitos dos fármacos , Proteínas de Ciclo Celular/metabolismo , Dopamina/metabolismo , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Células PC12 , Ligação Proteica , Ratos , Transdução de Sinais
15.
PLoS Negl Trop Dis ; 15(2): e0009105, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33544727

RESUMO

The argasid tick Ornithodoros moubata is the main vector of human relapsing fever (HRF) and African swine fever (ASF) in Africa. Salivary proteins are part of the host-tick interface and play vital roles in the tick feeding process and the host infection by tick-borne pathogens; they represent interesting targets for immune interventions aimed at tick control. The present work describes the transcriptome profile of salivary glands of O. moubata and assesses the gene expression dynamics along the trophogonic cycle using Illumina sequencing. De novo transcriptome assembling resulted in 71,194 transcript clusters and 41,011 annotated transcripts, which represent 57.6% of the annotation success. Most salivary gene expression takes place during the first 7 days after feeding (6,287 upregulated transcripts), while a minority of genes (203 upregulated transcripts) are differentially expressed between 7 and 14 days after feeding. The functional protein groups more abundantly overrepresented after blood feeding were lipocalins, proteases (especially metalloproteases), protease inhibitors including the Kunitz/BPTI-family, proteins with phospholipase A2 activity, acid tail proteins, basic tail proteins, vitellogenins, the 7DB family and proteins involved in tick immunity and defence. The complexity and functional redundancy observed in the sialotranscriptome of O. moubata are comparable to those of the sialomes of other argasid and ixodid ticks. This transcriptome provides a valuable reference database for ongoing proteomics studies of the salivary glands and saliva of O. moubata aimed at confirming and expanding previous data on the O. moubata sialoproteome.


Assuntos
Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Ornithodoros/genética , Ornithodoros/metabolismo , Transcriptoma , África , Febre Suína Africana , Animais , Asfarviridae , Feminino , Expressão Gênica , Imunidade , Ixodidae/genética , Ixodidae/metabolismo , Redes e Vias Metabólicas/genética , Ornithodoros/imunologia , Ornithodoros/virologia , Fosfolipases A2/metabolismo , Proteômica/métodos , Saliva , Glândulas Salivares , Suínos
16.
Artigo em Inglês | MEDLINE | ID: mdl-33548504

RESUMO

Scylla paramamosain is an economically important cultured crab species in China. Cyclins and cyclin-dependent kinases (CDKs) play important roles in regulations of cell cycle and ovarian development. MiRNAs can negatively regulate gene expression at the post-transcriptional level through base-complementary pairing with the 3'-untranslated region (3-UTR) of the target gene. In this study, bioinformatics prediction showed that miR-9c and miR-263a identified from our group's gonad miRNAome of S. paramamosain may bind to the 3' UTR region of cyclin A, cyclin B, cyclin E, cyclin H, CDK1, and CDK2. Furthermore, the results of double luciferase reporter gene assay showed that the luciferase activities of HEK293T cells co-transfected with miR-9c mimics/miR-9c inhibitor and the 3'-UTR plasmid vectors of the five genes (cyclin A, cyclin B, cyclin H, CDK1, and CDK2) were significantly decreased/increased compared with those in the NC (negative control) and BC (blank control) groups. The results in miR-263a were similar to miR-9c, but all of the six genes could be regulated by miR-263a. In in vivo experiments, agomiR-9c (miR-9c enhancer) injection resulted in decreases of cyclin A and CDK1 expression level, and reverse effects were observed by injecting antagomiR-9c. AgomiR-263a decreased the expression of cyclin A, cyclin B, cyclin H, CDK1, and CDK2, but antagomiR-263a increased their expression. Both the in vitro and in vivo experiments confirmed functions of miR-9c and miR-263a in cell cycle progress of ovarian development by expression regulation of cyclin A, cyclin B, cyclin E, cyclin H, CDK1, and CDK2. The findings provide new insights into the reproductive regulation mechanism in mud crab and further enrich the knowledge of cell cycle and ovarian development regulation in invertebrates.


Assuntos
Proteínas de Artrópodes/metabolismo , Braquiúros/metabolismo , Ciclinas/metabolismo , MicroRNAs/metabolismo , Ovário/metabolismo , Animais , Proteínas de Artrópodes/genética , Braquiúros/genética , Ciclinas/genética , Feminino , MicroRNAs/genética
17.
Ticks Tick Borne Dis ; 12(3): 101677, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33549977

RESUMO

Anaplasma ovis, a tick-borne intra-erythrocytic Gram-negative bacterium, is a causative agent of ovine anaplasmosis. It is known that Dermacentor ticks act as biological vectors for A. ovis. VirD4 is the machine component of Type IV Secretion System of A. ovis. To better understand the pathogen-vector interaction, VirD4 was used as a bait protein for screening midgut proteins of Dermacentor silvarum via yeast two-hybrid mating assay. As a result, a ribosomal protein RL12 was identified from the midgut cDNA library of D. silvarum. For further validation, using in vitro Glutathione S-transferase (GST) pull-down assay, interaction between the proteins, GST-RL12 and HIS-VirD4, was observed in Western blot analysis. The study is first of its kind reporting a D. silvarum midgut protein interaction with VirD4 from A. ovis. Functional annotations showed some important cellular processes are attributed to the protein, particularly in the stringent response and biogenesis. The results of the study suggest the involvement of the VirD4-RL12 interaction in the regulation of signaling pathways, which is a tool for understanding the pathogen-vector interaction.


Assuntos
Anaplasma ovis/genética , Vetores Aracnídeos/genética , Proteínas de Artrópodes/genética , Proteínas de Bactérias/genética , Dermacentor/genética , Proteínas Ribossômicas/genética , Anaplasma ovis/metabolismo , Animais , Vetores Aracnídeos/metabolismo , Vetores Aracnídeos/microbiologia , Proteínas de Artrópodes/metabolismo , Proteínas de Bactérias/metabolismo , Dermacentor/metabolismo , Dermacentor/microbiologia , Sistema Digestório/metabolismo , Sistema Digestório/microbiologia , Proteínas Ribossômicas/metabolismo
18.
J Immunol ; 206(6): 1140-1150, 2021 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-33526439

RESUMO

Intestinal microbiota are closely related to host physiology. Over the long course of evolution and interaction, both commensal bacteria and their host have evolved multiple strategies to adapt to each other. However, in invertebrates, the regulatory mechanism of intestinal microbiota homeostasis is largely unknown. In the current study, a digestive tract-specific C-type lectin, designated as CTL33, was identified because of its abundance and response to bacteria in the intestine of kuruma shrimp (Marsupenaeus japonicus). Silencing of CTL33 expression led directly to intestinal dysbiosis, tissue damage, and shrimp death. CTL33 could facilitate biofilm formation by the intestinal bacteria. This function originated from its unique architecture, with a lectin domain responsible for bacteria recognition and a coiled coil region that mediated CTL33 dimerization and cross-linked the bacteria into a biofilm-like complex. By mediating the formation of a biofilm, CTL33 promoted the establishment of intestinal bacteria in intestine and maintained the homeostasis of the microbiota. Thus, to our knowledge, we demonstrated a new mechanism of C-type lectin-mediated biofilm formation by intestinal bacteria, providing new insights into intestinal homeostasis regulation in invertebrates.


Assuntos
Proteínas de Artrópodes/metabolismo , Bactérias/imunologia , Microbioma Gastrointestinal/imunologia , Lectinas Tipo C/metabolismo , Penaeidae/imunologia , Animais , Proteínas de Artrópodes/genética , Biofilmes , Disbiose/genética , Disbiose/imunologia , Disbiose/microbiologia , Técnicas de Silenciamento de Genes , Homeostase/imunologia , Interações entre Hospedeiro e Microrganismos/imunologia , Lectinas Tipo C/genética , Penaeidae/metabolismo , Penaeidae/microbiologia , Domínios Proteicos
19.
Sci Rep ; 11(1): 921, 2021 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-33441720

RESUMO

House dust mites (HDMs) are one of the major causes of allergies in the world. The group 23 allergen, Der p 23, from Dermatophagoides pteronyssinus, is a major allergen amongst HDM-sensitized individuals. This study aims to determine the specific immunoglobulin E (sIgE) binding frequency and IgE-binding residues of recombinant Der p 23 (rDer p 23) allergen amongst a cohort of consecutive atopic individuals in a tropical region. We performed site-directed mutagenesis and carried out immuno-dot blot assays using 65 atopic sera. The immuno-dot blot assays results indicated that the two residues K44 and E46 which are located at the N-terminal region are the major IgE-binding residues. The rDerp-23 sIgE titers are strongly correlated to the number of IgE-binding residues for rDer p 23 (P < 0.001). Atopic individuals who were only sensitized to HDM have a significantly higher number of IgE-binding residues than the individuals who were polysensitized to HDM and other crude allergens (P < 0.05). Individuals with allergic multimorbidity and moderate-to-severe allergic rhinitis also have a higher number of IgE-binding residues compared to those with single allergic disease and mild allergic rhinitis. The results prompt us to hypothesize that the individuals who have a higher number of IgE-binding residues may face a bigger challenge to be treated through immunotherapy due to the complexity in designing an effective hypoallergen with a high number of IgE-binding residues. We propose that the development of a refined molecular diagnostic assay, which includes alanine substitution of surface-exposed residues could be a more precise diagnostic strategy to identify all the IgE-binding residues of a major allergen for an atopic individual and the development could be another new dimension in allergy diagnosis and allergen immunotherapy treatment.


Assuntos
Antígenos de Dermatophagoides/imunologia , Hipersensibilidade/imunologia , Pyroglyphidae/imunologia , Adulto , Alérgenos/imunologia , Animais , Antígenos de Dermatophagoides/metabolismo , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/metabolismo , Dermatophagoides pteronyssinus/imunologia , Poeira/imunologia , Feminino , Humanos , Hipersensibilidade Imediata , Immunoblotting , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina E/metabolismo , Masculino , Rinite Alérgica , Adulto Jovem
20.
Int J Biol Macromol ; 174: 207-215, 2021 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-33482212

RESUMO

Phenoloxidase (PO) is a typical metal enzyme, which requires metal ions as prosthetic groups to enable the full exertion of its activity. To study how metal ions affected the activity and structure of PO enzymes, while providing reference materials for in-depth investigations, we examined the effects of different metal ions (Cu2+, Zn2+, Mg2+, Ca2+, and Ba2+) on their activities. Furthermore, Cu2+ and Mg2+ were selected for further investigation through UV spectra, intrinsic fluorescence spectroscopy, AFM, and FTIR. It was revealed that Cu2+ had a more obvious effect on PO compared to Mg2+. The PO could be activated when the concentrations of Cu2+ and Mg2+ were lower than 10-3 and 10-2 mol/L, respectively, and maximum PO activities (182.14% and 141.02%) were observed at 10-4 mol/L concentrations of Cu2+ and Mg2+. When the concentrations of Cu2+ and Mg2+ were higher than 10-2 and 10-1 mol/L, the activities PO were inhibited. The results of the UV-vis and fluorescence spectra revealed that Cu2+ shaped the tertiary structure of PO, whereas the effect of Mg2+ was slight. The AFM results demonstrated that high concentrations of Cu2+ and Mg2+ resulted in PO aggregation. FTIR analysis indicated that the total content of PO α-helices and ß-sheets decreased with higher concentrations of Cu2+ and Mg2+.


Assuntos
Cobre/farmacologia , Magnésio/farmacologia , Monofenol Mono-Oxigenase/química , Monofenol Mono-Oxigenase/metabolismo , Penaeidae/enzimologia , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Transferência Ressonante de Energia de Fluorescência , Microscopia de Força Atômica , Estrutura Secundária de Proteína/efeitos dos fármacos , Estrutura Terciária de Proteína/efeitos dos fármacos , Espectrometria de Fluorescência
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