ABSTRACT
The Sox family of transcription factors regulates many processes during metazoan development, including stem cell maintenance and nervous system specification. Characterizing the repertoires and roles of these genes can therefore provide important insights into animal evolution and development. We further characterized the Sox repertoires of several arachnid species with and without an ancestral whole-genome duplication and compared their expression between the spider Parasteatoda tepidariorum and the harvestman Phalangium opilio. We found that most Sox families have been retained as ohnologs after whole-genome duplication and evidence for potential subfunctionalization and/or neofunctionalization events. Our results also suggest that Sox21b-1 likely regulated segmentation ancestrally in arachnids, playing a similar role to the closely related SoxB gene, Dichaete, in insects. We previously showed that Sox21b-1 is required for the simultaneous formation of prosomal segments and sequential addition of opisthosomal segments in P. tepidariorum. We studied the expression and function of Sox21b-1 further in this spider and found that although this gene regulates the generation of both prosomal and opisthosomal segments, it plays different roles in the formation of these tagmata reflecting their contrasting modes of segmentation and deployment of gene regulatory networks with different architectures.
Subject(s)
Arachnida/genetics , Evolution, Molecular , SOX Transcription Factors/genetics , Animals , Arachnida/embryology , Arachnida/metabolism , Female , Male , SOX Transcription Factors/metabolismABSTRACT
Hemocyanin (Hc), a copper-containing extracellular multimeric protein, is the major protein component of hemolymph in different arachnid groups. Hc possesses 7 or 8 very well-characterized types of monomers with molecular weights ranging from 70 to 85 kDa, organized in hexamers or multiple of hexamers. The present chapter compiles the existing data with relation to the function of this protein in the arachnids. Hc has as main function the reversible transport of O2, but it shows many secondary though not less important functions. With reference to this, it has been described that Hc can transport hydrophobic molecules (lipid-derived hormones and lipids) to the different organs, having a key role in the lipid transport system. In arachnids, like in other arthropods and invertebrates, Hc has phenoloxidase function which is related to different metabolic processes such as melanin formation and defense against pathogens. In addition, Hc has additional defensive functions since it can serve as precursor for the production of antimicrobial peptides. In short, the evolution of this protein has led to the development of multiple functions essential for organisms possessing this protein.
Subject(s)
Arachnida , Hemocyanins , Animals , Arachnida/enzymology , Arachnida/metabolism , Hemocyanins/metabolism , Monophenol Monooxygenase/metabolismABSTRACT
Despite application of genome-scale datasets, the phylogenetic placement of scorpions within arachnids remains contentious between two different phylogenetic data classes. Paleontologists continue to recover scorpions in a basally branching position, partly owing to their morphological similarity to extinct marine orders like Eurypterida (sea scorpions). Phylogenomic datasets consistently recover scorpions in a derived position, as the sister group of Tetrapulmonata (a clade of arachnids that includes spiders). To adjudicate between these hypotheses using a rare genomic change (RGC), we leveraged the recent discovery of ancient paralogy in spiders and scorpions to assess phylogenetic placement. We identified homologs of four transcription factors required for appendage patterning (dachshund, homothorax, extradenticle, and optomotor blind) in arthropods that are known to be duplicated in spiders. Using genomic resources for a spider, a scorpion, and a harvestman, we conducted gene tree analyses and assayed expression patterns of scorpion gene duplicates. Here we show that scorpions, like spiders, retain two copies of all four transcription factors, whereas arachnid orders like mites and harvestmen bear a single copy. A survey of embryonic expression patterns of the scorpion paralogs closely matches those of their spider counterparts, with one paralog consistently retaining the putatively ancestral pattern found in the harvestman, as well as the mite, and/or other outgroups. These data comprise a rare genomic change in chelicerate phylogeny supporting the inference of a distal placement of scorpions. Beyond demonstrating the diagnostic power of developmental genetic data as a phylogenetic data class, a derived placement of scorpions within the arachnids, together with an array of stem-group Paleozoic scorpions that occupied marine habitats, effectively rules out a scenario of a single colonization of terrestrial habitat within Chelicerata, even in tree topologies contrived to recover the monophyly of Arachnida.
Subject(s)
Arachnida/classification , Arachnida/genetics , Homeodomain Proteins/genetics , Nerve Tissue Proteins/genetics , T-Box Domain Proteins/genetics , Transcription Factors/genetics , Animals , Arachnida/embryology , Arachnida/metabolism , Female , Gene Dosage , Gene Expression Regulation, Developmental , Genes, Developmental/genetics , Genomics , Mites/genetics , Phylogeny , Scorpions/embryology , Scorpions/genetics , Scorpions/metabolism , Spiders/embryology , Spiders/genetics , Spiders/metabolismABSTRACT
Dragline silk has the highest tensile strength among the seven types of spider silks due to its abundant polyalanine motifs. Whereas the flagelliform spider silk is most extensible as its composed spidroin is rich in GPGGX motifs. Most of the spider silk proteins are composed of an extensive repetitive domain flanked by N- and C-terminal domains. To obtain artificial fibers with considerable strength and extensibility, herein a kind of chimeric minispidroins were constructed whose repetitive domain (R) mainly consisted of polyalanine motifs and GPGGX motifs. In our study, NT and CT from Araneus ventricosus MaSp1 were fused with different numbers (1, 4, 8) of repeat domains (R), resulting in three chimeric minispidroins. All these chimeric proteins could form silk-like fibers via manual pulling. As the chimeric spidroin was pulled from the protein solution into fiber by shear forces, the secondary structure transformed from α-helix to ß-sheet. Among the three types of fibers, the average tensile strength of NTR4CT ranked the highest (149 MPa), which could provide outstanding material with better mechanical properties. In addition, NT was fused with CT and repetitive domain R respectively, namely NC and NR proteins. As a result, NC could form fibers that had much lower properties than NTR1CT, indicating that repetitive domain was responsible for the strength and elasticity of the fibers. However, NR did not form silk-like fibers, suggesting that Araneus ventricosus Masp1 CT controlled fiber formation. These results broaden the limited knowledge of chimeric spider silk sequences.
Subject(s)
Arachnida/metabolism , Fibroins/chemistry , Insect Proteins/chemistry , Proteins/chemistry , Recombinant Fusion Proteins/chemistry , Animals , Cloning, Molecular , Elasticity , Escherichia coli/genetics , Protein Structure, Secondary , Tensile StrengthABSTRACT
Myosin II molecules in the thick filaments of striated muscle form a structure in which the heads interact with each other and fold back onto the tail. This structure, the "interacting heads motif" (IHM), provides a mechanistic basis for the auto-inhibition of myosin in relaxed thick filaments. Similar IHM interactions occur in single myosin molecules of smooth and nonmuscle cells in the switched-off state. In addition to the interaction between the two heads, which inhibits their activity, the IHM also contains an interaction between the motor domain of one head and the initial part (subfragment 2, S2) of the tail. This is thought to be a crucial anchoring interaction that holds the IHM in place on the thick filament. S2 appears to cross the head at a specific location within a broader region of the motor domain known as the myosin mesa. Here, we show that the positive and negative charge distribution in this part of the mesa is complementary to the charge distribution on S2. We have designated this the "mesa trail" owing to its linear path across the mesa. We studied the structural sequence alignment, the location of charged residues on the surface of myosin head atomic models, and the distribution of surface charge potential along the mesa trail in different types of myosin II and in different species. The charge distribution in both the mesa trail and the adjacent S2 is relatively conserved. This suggests a common basis for IHM formation across different myosin IIs, dependent on attraction between complementary charged patches on S2 and the myosin head. Conservation from mammals to insects suggests that the mesa trail/S2 interaction plays a key role in the inhibitory function of the IHM.
Subject(s)
Myosin Type II/metabolism , Animals , Arachnida/chemistry , Arachnida/metabolism , Arthropod Proteins/chemistry , Arthropod Proteins/metabolism , Dictyostelium/chemistry , Dictyostelium/metabolism , Insecta , Mammals , Models, Molecular , Myosin Type II/chemistry , Protein Interaction Domains and Motifs , Protozoan Proteins/chemistry , Protozoan Proteins/metabolism , Species SpecificityABSTRACT
Diverse fields of modern technology and biomedicine can benefit from the application of ready-to-use chitin-based scaffolds. In this work we show for the first time the applicability of tubular and porous chitin from Caribena versicolor spiders as a scaffold for the development of an effective CuO/Cu(OH)2 catalyst for the reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AM), and as a scaffold for the tissue engineering of selected cells. The formation of CuO/Cu(OH)2 phases on and within the chitinous tubes leads to a hybrid material with excellent catalytic performance with respect to the reduction of p-nitrophenol. On the other hand, experimental results provide for the first time strong evidence for the biocompatibility of spider chitin with different cell types, a human progenitor cell line (hPheo1), as well as cardiomyocytes differentiated from induced pluripotent stem cells (iPSC-CMs) that were cultured on a tube-like scaffold.
Subject(s)
Arachnida/metabolism , Biomimetic Materials/chemistry , Chitin/chemistry , Tissue Engineering , Tissue Scaffolds/chemistry , Animals , Cells, Cultured , HumansABSTRACT
Spider silk is a biomaterial with exceptional mechanical toughness, and there is great interest in developing biomimetic methods to produce engineered spider silk-based materials. However, the mechanisms that regulate the conversion of spider silk proteins (spidroins) from highly soluble dope into silk are not completely understood. The N-terminal domain (NT) of Euprosthenops australis dragline silk protein undergoes conformational and quaternary-structure changes from a monomer at a pH above 7 to a homodimer at lower pH values. Conversion from the monomer to the dimer requires the protonation of three conserved glutamic acid residues, resulting in a low-pH `locked' dimer stabilized by symmetric electrostatic interactions at the poles of the dimer. The detailed molecular events during this transition are still unresolved. Here, a 2.1â Å resolution crystal structure of an NT T61A mutant in an alternative, asymmetric, dimer form in which the electrostatic interactions at one of the poles are dramatically different from those in symmetrical dimers is presented. A similar asymmetric dimer structure from dragline silk of Nephila clavipes has previously been described. It is suggested that asymmetric dimers represent a conserved intermediate state in spider silk formation, and a revised `lock-and-trigger' mechanism for spider silk formation is presented.
Subject(s)
Arachnida/metabolism , Fibroins/chemistry , Recombinant Proteins/chemistry , Animals , Crystallization/methods , Escherichia coli/genetics , Fibroins/genetics , Models, Molecular , Molecular Structure , Mutation , Protein Domains , Protein Multimerization , Protein Structure, Quaternary , Static ElectricityABSTRACT
In silico and in vitro studies have made progress in understanding protein-protein complex formation; however, the molecular mechanisms for their dissociation are unclear. Protein-protein complexes, lasting from microseconds to years, often involve induced-fit, challenging computational or kinetic analysis. Charybdotoxin (CTX), a peptide from the Leiurus scorpion venom, blocks voltage-gated K+-channels in a unique example of binding/unbinding simplicity. CTX plugs the external mouth of K+-channels pore, stopping K+-ion conduction, without inducing conformational changes. Conflicting with a tight binding, we show that external permeant ions enhance CTX-dissociation, implying a path connecting the pore, in the toxin-bound channel, with the external solution. This sensitivity is explained if CTX wobbles between several bound conformations, producing transient events that restore the electrical and ionic trans-pore gradients. Wobbling may originate from a network of contacts in the interaction interface that are in dynamic stochastic equilibria. These partially-bound intermediates could lead to distinct, and potentially manipulable, dissociation pathways.
Subject(s)
Charybdotoxin/metabolism , Ions/metabolism , Potassium Channels, Voltage-Gated/chemistry , Potassium Channels, Voltage-Gated/metabolism , Potassium/metabolism , Animals , Arachnida/metabolism , Potassium Channels, Voltage-Gated/drug effects , Protein Binding , Protein ConformationABSTRACT
The roles of surrounding membrane lipids in the functions of transmembrane and peripheral membrane proteins are largely unknown. Herein, we utilize the recently reported structures of the TRPV1 ion channel protein bound to its potent protein agonist, the double-knot toxin (DkTx), as a model system to investigate the roles of toxin-lipid interfaces in TRPV1 activation by characterizing a series of DkTx variants electrophysiologically. Together with membrane partitioning experiments, these studies reveal that toxin-lipid interfaces play an overwhelmingly dominant role in channel activation as compared to lipid-devoid toxin-channel interfaces. Additionally, we find that whereas the membrane interfaces formed by one of the knots of the toxin endow it with its low channel-dissociation rate, those formed by other knot contribute primarily to its potency. These studies establish that protein-lipid interfaces play nuanced yet profound roles in the function of protein-protein complexes within membranes.
Subject(s)
Arachnida/metabolism , Arthropod Proteins/metabolism , Membrane Lipids/metabolism , TRPV Cation Channels/agonists , TRPV Cation Channels/metabolism , Animals , Models, Molecular , RatsABSTRACT
Homeobox genes are key toolkit genes that regulate the development of metazoans and changes in their regulation and copy number have contributed to the evolution of phenotypic diversity. We recently identified a whole genome duplication (WGD) event that occurred in an ancestor of spiders and scorpions (Arachnopulmonata), and that many homeobox genes, including two Hox clusters, appear to have been retained in arachnopulmonates. To better understand the consequences of this ancient WGD and the evolution of arachnid homeobox genes, we have characterized and compared the homeobox repertoires in a range of arachnids. We found that many families and clusters of these genes are duplicated in all studied arachnopulmonates (Parasteatoda tepidariorum, Pholcus phalangioides, Centruroides sculpturatus, and Mesobuthus martensii) compared with nonarachnopulmonate arachnids (Phalangium opilio, Neobisium carcinoides, Hesperochernes sp., and Ixodes scapularis). To assess divergence in the roles of homeobox ohnologs, we analyzed the expression of P. tepidariorum homeobox genes during embryogenesis and found pervasive changes in the level and timing of their expression. Furthermore, we compared the spatial expression of a subset of P. tepidariorum ohnologs with their single copy orthologs in P. opilio embryos. We found evidence for likely subfunctionlization and neofunctionalization of these genes in the spider. Overall our results show a high level of retention of homeobox genes in spiders and scorpions post-WGD, which is likely to have made a major contribution to their developmental evolution and diversification through pervasive subfunctionlization and neofunctionalization, and paralleling the outcomes of WGD in vertebrates.
Subject(s)
Arachnida/genetics , Evolution, Molecular , Gene Duplication , Genes, Homeobox , Animals , Arachnida/embryology , Arachnida/metabolism , Embryonic Development , Gene Expression , Multigene Family , TranscriptomeABSTRACT
Both CC and CXC-class chemokines drive inflammatory disease. Tick salivary chemokine-binding proteins (CKBPs), or evasins, specifically bind subsets of CC- or CXC-chemokines, and could precisely target disease-relevant chemokines. Here we have used yeast surface display to identify two tick evasins: a CC-CKBP, P1243 from Amblyomma americanum and a CXC-CKBP, P1156 from Ixodes ricinus. P1243 binds 11 CC-chemokines with Kd < 10 nM, and 10 CC-chemokines with Kd between 10 and 100 nM. P1156 binds two ELR + CXC-chemokines with Kd < 10 nM, and four ELR + CXC-chemokines with Kd between 10 and 100 nM. Both CKBPs neutralize chemokine activity with IC50 < 10 nM in cell migration assays. As both CC- and CXC-CKBP activities are desirable in a single agent, we have engineered "two-warhead" CKBPs to create single agents that bind and neutralize subsets of CC and CXC chemokines. These results show that tick evasins can be linked to create non-natural proteins that target subsets of CC and CXC chemokines. We suggest that "two-warhead" evasins, designed by matching the activities of parental evasins to CC and CXC chemokines expressed in disease, would achieve precision targeting of inflammatory disease-relevant chemokines by a single agent.
Subject(s)
Receptors, CXCR/metabolism , Receptors, Chemokine/metabolism , Ticks/metabolism , Amino Acid Sequence , Animals , Arachnida/metabolism , Chemokines/metabolism , Chemokines, CC/metabolism , Chemokines, CXC/metabolism , Genetic Engineering , HEK293 Cells , Humans , Protein Binding , Receptors, CXCR/genetics , Saccharomyces cerevisiae/metabolism , THP-1 CellsABSTRACT
BACKGROUND: Pardosa pseudoannulata is a prevailing spider species, and has been regarded as an important bio-control agent of insect pests in farmland of China. However, the available genomic and transcriptomic databases of P. pseudoannulata and their venom are limited, which severely hampers functional genomic analysis of P. pseudoannulata. Recently high-throughput sequencing technology has been proved to be an efficient tool for profiling the transcriptome of relevant non-target organisms exposed to Bacillus thuringiensis (Bt) protein through food webs. RESULTS: In this study, the transcriptome of the venom apparatus was analyzed. A total of 113,358 non-redundant unigenes were yielded, among which 34,041 unigenes with complete or various length encoding regions were assigned biological function annotations and annotated with gene ontology and karyotic orthologous group terms. In addition, 3726 unigenes involved in response to stimulus and 720 unigenes associated with immune-response pathways were identified. Furthermore, we investigated transcriptomic changes in the venom apparatus using tag-based DGE technique. A total of 1724 differentially expressed genes (DEGs) were detected, while 75 and 372 DEGs were functionally annotated with KEGG pathways and GO terms, respectively. qPCR analyses were performed to verify the DEGs directly or indirectly related to immune and stress responses, including genes encoding heat shock protein, toll-like receptor, GST and NADH dehydrogenase. CONCLUSION: This is the first study conducted to specifically investigate the venom apparatus of P. pseudoannulata in response to Bt protein exposure through tritrophic chain. A substantial fraction of transcript sequences was generated by high-throughput sequencing of the venom apparatus of P. pseudoannulata. Then a comparative transcriptome analysis showing a large number of candidate genes involved in immune response were identified by the tag-based DGE technology. This transcriptome dataset will provide a comprehensive sequence resource for furture molecular genetic research of the venom apparatus of P. pseudoannulata.
Subject(s)
Arachnida/genetics , Bacterial Proteins/pharmacology , Endotoxins/pharmacology , Genes, Insect/genetics , Hemolysin Proteins/pharmacology , Spider Venoms/genetics , Transcriptome/genetics , Animals , Arachnida/drug effects , Arachnida/metabolism , Arachnida/physiology , Bacillus thuringiensis Toxins , Gene Expression Regulation/drug effects , Gene Expression Regulation/genetics , High-Throughput Nucleotide Sequencing , RNA, Messenger/analysis , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Analysis, DNA , Spider Venoms/analysis , Spider Venoms/metabolism , Transcriptome/drug effectsABSTRACT
BACKGROUND: Stegodyphus mimosarum is a candidate model organism belonging to the class Arachnida in the phylum Arthropoda. Studies on the biology of S. mimosarum over the past several decades have consisted of behavioral research and comparison of gene sequences based on the assembled genome sequence. Given the lack of systematic protein analyses and the rich source of information in the genome, we predicted the relationships of proteins in S. mimosarum by bioinformatics comparison with genome-wide proteins from select model organisms using gene mapping. RESULTS: The protein-protein interactions (PPIs) of 11 organisms were integrated from four databases (BioGrid, InAct, MINT, and DIP). Here, we present comprehensive prediction and analysis of 3810 proteins in S. mimosarum with regard to interactions between proteins using PPI data of organisms. Interestingly, a portion of the protein interactions conserved among Saccharomyces cerevisiae, Homo sapiens, Arabidopsis thaliana, and Drosophila melanogaster were found to be associated with RNA splicing. In addition, overlap of predicted PPIs in reference organisms, Gene Ontology, and topology models in S. mimosarum are also reported. CONCLUSIONS: Addition of Stegodyphus, a spider representative of interactomic research, provides the possibility of obtaining deeper insights into the evolution of PPI networks among different animal species. This work largely supports the utility of the "stratus clouds" model for predicted PPIs, providing a roadmap for integrative systems biology in S. mimosarum.
Subject(s)
Arachnida/metabolism , Arthropod Proteins/metabolism , Protein Interaction Mapping , Animals , Arachnida/genetics , Arthropod Proteins/genetics , Databases, Protein , Gene Ontology , RNA Splicing , Species SpecificityABSTRACT
Lepidopteran silks number in the thousands and display a vast diversity of structures, properties and industrial potential. To map this remarkable biochemical diversity, we present an identification and screening method based on the infrared spectra of native silk feedstock and cocoons. Multivariate analysis of over 1214 infrared spectra obtained from 35 species allowed us to group silks into distinct hierarchies and a classification that agrees well with current phylogenetic data and taxonomies. This approach also provides information on the relative content of sericin, calcium oxalate, phenolic compounds, poly-alanine and poly(alanine-glycine) ß-sheets. It emerged that the domesticated mulberry silkmoth Bombyx mori represents an outlier compared with other silkmoth taxa in terms of spectral properties. Interestingly, Epiphora bauhiniae was found to contain the highest amount of ß-sheets reported to date for any wild silkmoth. We conclude that our approach provides a new route to determine cocoon chemical composition and in turn a novel, biological as well as material, classification of silks.
Subject(s)
Arachnida/metabolism , Moths/metabolism , Silk/chemistry , Silk/classification , Animals , Bombyx/metabolism , Calcium Oxalate/chemistry , Peptides/chemistry , Phenols/chemistry , Phylogeny , Sericins/chemistry , Spectrophotometry, InfraredABSTRACT
Phosphorylation of myosin regulatory light chain (RLC) N-terminal extension (NTE) activates myosin in thick filaments. RLC phosphorylation plays a primary regulatory role in smooth muscles and a secondary (modulatory) role in striated muscles, which is regulated by Ca(2+)via TnC/TM on the thin filament. Tarantula striated muscle exhibits both regulatory systems: one switches on/off contraction through thin filament regulation, and another through PKC constitutively Ser35 phosphorylated swaying free heads in the thick filaments that produces quick force on twitches regulated from 0 to 50% and modulation is accomplished recruiting additional force-potentiating free and blocked heads via Ca(2+)4-CaM-MLCK Ser45 phosphorylation. We have used microsecond molecular dynamics (MD) simulations of tarantula RLC NTE to understand the structural basis for phosphorylation-based regulation in tarantula thick filament activation. Trajectory analysis revealed that an inter-domain salt bridge network (R39/E58,E61) facilitates the formation of a stable helix-coil-helix (HCH) motif formed by helices P and A in the unphosphorylated NTE of both myosin heads. Phosphorylation of the blocked head on Ser45 does not induce any substantial structural changes. However, phosphorylation of the free head on Ser35 disrupts this salt bridge network and induces a partial extension of helix P along RLC helix A. While not directly participating in the HCH folding, phosphorylation of Ser35 unlocks a compact structure and allows the NTE to spontaneously undergo coil-helix transitions. The modest structural change induced by the subsequent Ser45 diphosphorylation monophosphorylated Ser35 free head facilitates full helix P extension into a single structurally stable α-helix through a network of intra-domain salt bridges (pS35/R38,R39,R42). We conclude that tarantula thick filament activation is controlled by sequential Ser35-Ser45 phosphorylation via a conserved disorder-to-order transition.
Subject(s)
Actin Cytoskeleton/chemistry , Muscle, Skeletal/chemistry , Muscle, Smooth/metabolism , Myosin Light Chains/chemistry , Actin Cytoskeleton/metabolism , Animals , Arachnida/chemistry , Arachnida/metabolism , Calcium/metabolism , Molecular Dynamics Simulation , Muscle, Skeletal/metabolism , Muscle, Smooth/chemistry , Myosin Light Chains/metabolism , Phosphorylation , Protein Kinase C/chemistry , Protein Structure, SecondaryABSTRACT
Molecular dynamics simulations of smooth and striated muscle myosin regulatory light chain (RLC) N-terminal extension (NTE) showed that diphosphorylation induces a disorder-to-order transition. Our goal here was to further explore the effects of mono- and diphosphorylation on the straightening and rigidification of the tarantula myosin RLC NTE. For that we used MD simulations followed by persistence length analysis to explore the consequences of secondary and tertiary structure changes occurring on RLC NTE following phosphorylation. Static and dynamic persistence length analysis of tarantula RLC NTE peptides suggest that diphosphorylation produces an important 24-fold straightening and a 16-fold rigidification of the RLC NTE, while monophosphorylation has a less profound effect. This new information on myosin structural mechanics, not fully revealed by previous EM and MD studies, add support to a cooperative phosphorylation-dependent activation mechanism as proposed for the tarantula thick filament. Our results suggest that the RLC NTE straightening and rigidification after Ser45 phosphorylation leads to a release of the constitutively Ser35 monophosphorylated free head swaying away from the thick filament shaft. This is so because the stiffened diphosphorylated RLC NTE would hinder the docking back of the free head after swaying away, becoming released and mobile and unable to recover its original interacting position on activation.
Subject(s)
Muscle, Smooth/chemistry , Myosin Light Chains/chemistry , Protein Folding , Animals , Arachnida/chemistry , Arachnida/metabolism , Calcium/metabolism , Molecular Dynamics Simulation , Muscle, Smooth/metabolism , Myosin Light Chains/metabolism , Phosphorylation , Protein Structure, SecondaryABSTRACT
The public deposition of large transcriptome shotgun assembly (TSA) datasets for the Araneae (true spiders) provides a resource for determining the structures of the native neuropeptides present in members of this chelicerate order. Here, the Araneae TSA data were mined for putative peptide-encoding transcripts using the recently deduced neuropeptide precursors from the Western black widow Latrodectus hesperus as query templates. Neuropeptide-encoding transcripts from five spiders, Latrodectus tredecimguttatus, Stegodyphus mimosarum, Stegodyphus lineatus, Stegodyphus tentoriicola and Acanthoscurria geniculata, were identified, including ones encoding members of the allatostatin A, allatostatin B, allatostatin C, allatotropin, CAPA/periviscerokinin/pyrokinin, crustacean cardioactive peptide, crustacean hyperglycemic hormone/ion transport peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FMRFamide-like peptide (FLP), GSEFLamide, insulin-like peptide, orcokinin, proctolin, short neuropeptide F, SIFamide, sulfakinin and tachykinin-related peptide (TRP) families. A total of 156 distinct peptides were predicted from the precursor proteins deduced from the S. mimosarum transcripts, with 65, 26, 21 and 12 peptides predicted from those deduced from the A. geniculata, L. tredecimguttatus, S. lineatus and S. tentoriicola sequences, respectively. Among the peptides identified were variant isoforms of FLP, orcokinin and TRP, peptides whose structures are similar to ones previously identified from L. hesperus. The prediction of these atypical peptides from multiple spiders suggests that they may be broadly conserved within the Araneae rather than being species-specific variants. Taken collectively, the data described here greatly expand the number of known Araneae neuropeptides, providing a foundation for future functional studies of peptidergic signaling in this important Chelicerate order.
Subject(s)
Arachnida/metabolism , Black Widow Spider/metabolism , Computational Biology/methods , Neuropeptides/classification , Neuropeptides/metabolism , Amino Acid Sequence , Animals , Arachnida/classification , Molecular Sequence Data , Neuropeptides/analysis , Reference StandardsABSTRACT
Opiliones are one of the largest arachnid orders, with more than 6,500 species in 50 families. Many of these families have been erected or reorganized in the last few years since the publication of The Biology of Opiliones. Recent years have also seen an explosion in phylogenetic work on Opiliones, as well as in studies using Opiliones as test cases to address biogeographic and evolutionary questions more broadly. Accelerated activity in the study of Opiliones evolution has been facilitated by the discovery of several key fossils, including the oldest known Opiliones fossil, which represents a new, extinct suborder. Study of the group's biology has also benefited from rapid accrual of genomic resources, particularly with respect to transcriptomes and functional genetic tools. The rapid emergence and utility of Phalangium opilio as a model for evolutionary developmental biology of arthropods serve as demonstrative evidence of a new area of study in Opiliones biology, made possible through transcriptomic data.
Subject(s)
Arachnida/physiology , Biological Evolution , Animals , Arachnida/genetics , Arachnida/growth & development , Arachnida/metabolism , Biodiversity , Fossils , Genome , Models, Animal , Phylogeny , TranscriptomeABSTRACT
Arthropods produce a great variety of natural compounds, many of which have unexplored biosynthesis. Among the armored harvestmen (Arachnida: Opiliones) of the suborder Laniatores, the defensive gland exudates contain vinyl ketones and other constituents of supposed polyketide origin. We have studied the biosynthesis of 1-hepten-3-one in the Neotropical harvestman Iporangaia pustulosa by feeding individuals with ¹³C-labeled precursors, demonstrating its mixed acetate/propionate origin. ¹³C NMR spectroscopy showed an unusual labeling pattern suggesting different propionate sources for starting and extender units. Our analysis also indicates the presence of methylmalonyl-CoA mutase, converting acetate into propionyl-CoA via succinyl-CoA, together with other C3 unit routes. This is the first biosynthetic study of alkyl vinyl ketones in arthropods. Our results shed light on the origin and diversification of chemical compounds in a major arthropod group.
Subject(s)
Alkenes/metabolism , Arachnida/metabolism , Ketones/metabolism , Acyl Coenzyme A/metabolism , Animals , Arachnida/enzymology , Carbon Isotopes/chemistry , Ketones/chemistry , Magnetic Resonance Spectroscopy , Methylmalonyl-CoA Mutase/metabolism , Propionates/chemistry , Propionates/metabolismABSTRACT
Benzoquinones are usually present in arthropod defence exudates. Here, we describe the chemical profiles of 12 harvestman species belonging to the neotropical family Gonyleptidae. Nine of the studied species produced benzoquinones, while three produced alkyl phenols. Two benzoquinones and one phenol exhibited biological activity against bacteria and fungi. We also studied the biosynthesis of 2-ethyl-1,4-benzoquinone by feeding Magnispina neptunus individuals with ¹³C-labelled precursors; the benzoquinones were biosynthesised through a polyketide pathway using acetate and propionate building blocks.