Viral cystatin evolution and three-dimensional structure modelling: A case of directional selection acting on a viral protein involved in a host-parasitoid interaction

<p>Abstract</p> <p>Background</p> <p>In pathogens, certain genes encoding proteins that directly interact with host defences coevolve with their host and are subject to positive selection. In the lepidopteran host-wasp parasitoid system, one of the most original strategies developed by the wasps to defeat host defences is the injection of a symbiotic polydnavirus at the same time as the wasp eggs. The virus is essential for wasp parasitism success since viral gene expression alters the immune system and development of the host. As a wasp mutualist symbiont, the virus is expected to exhibit a reduction in genome complexity and evolve under wasp phyletic constraints. However, as a lepidopteran host pathogenic symbiont, the virus is likely undergoing strong selective pressures for the acquisition of new functions by gene acquisition or duplication. To understand the constraints imposed by this particular system on virus evolution, we studied a polydnavirus gene family encoding cyteine protease inhibitors of the cystatin superfamily.</p> <p>Results</p> <p>We show that <it>cystatins </it>are the first bracovirus genes proven to be subject to strong positive selection within a host-parasitoid system. A generated three-dimensional model of <it>Cotesia congregata </it>bracovirus cystatin 1 provides a powerful framework to position positively selected residues and reveal that they are concentrated in the vicinity of actives sites which interact with cysteine proteases directly. In addition, phylogenetic analyses reveal two different <it>cystatin </it>forms which evolved under different selective constraints and are characterized by independent adaptive duplication events.</p> <p>Conclusion</p> <p>Positive selection acts to maintain <it>cystatin </it>gene duplications and induces directional divergence presumably to ensure the presence of efficient and adapted cystatin forms. Directional selection has acted on key cystatin active sites, suggesting that cystatins coevolve with their host target. We can strongly suggest that cystatins constitute major virulence factors, as was already proposed in previous functional studies.</p

Significance of Cuscutain, a cysteine protease from Cuscuta reflexa, in host-parasite interactions

<p>Abstract</p> <p>Background</p> <p>Plant infestation with parasitic weeds like <it>Cuscuta reflexa </it>induces morphological as well as biochemical changes in the host and the parasite. These modifications could be caused by a change in protein or gene activity. Using a comparative macroarray approach <it>Cuscuta </it>genes specifically upregulated at the host attachment site were identified.</p> <p>Results</p> <p>One of the infestation specific <it>Cuscuta </it>genes encodes a cysteine protease. The protein and its intrinsic inhibitory peptide were heterologously expressed, purified and biochemically characterized. The haustoria specific enzyme was named cuscutain in accordance with similar proteins from other plants, e.g. papaya. The role of cuscutain and its inhibitor during the host parasite interaction was studied by external application of an inhibitor suspension, which induced a significant reduction of successful infection events.</p> <p>Conclusions</p> <p>The study provides new information about molecular events during the parasitic plant - host interaction. Inhibition of cuscutain cysteine proteinase could provide means for antagonizing parasitic plants.</p

The venom composition of the parasitic wasp Chelonus inanitus resolved by combined expressed sequence tags analysis and proteomic approach

<p>Abstract</p> <p>Background</p> <p>Parasitic wasps constitute one of the largest group of venomous animals. Although some physiological effects of their venoms are well documented, relatively little is known at the molecular level on the protein composition of these secretions. To identify the majority of the venom proteins of the endoparasitoid wasp <it>Chelonus inanitus </it>(Hymenoptera: Braconidae), we have randomly sequenced 2111 expressed sequence tags (ESTs) from a cDNA library of venom gland. In parallel, proteins from pure venom were separated by gel electrophoresis and individually submitted to a nano-LC-MS/MS analysis allowing comparison of peptides and ESTs sequences.</p> <p>Results</p> <p>About 60% of sequenced ESTs encoded proteins whose presence in venom was attested by mass spectrometry. Most of the remaining ESTs corresponded to gene products likely involved in the transcriptional and translational machinery of venom gland cells. In addition, a small number of transcripts were found to encode proteins that share sequence similarity with well-known venom constituents of social hymenopteran species, such as hyaluronidase-like proteins and an Allergen-5 protein.</p> <p>An overall number of 29 venom proteins could be identified through the combination of ESTs sequencing and proteomic analyses. The most highly redundant set of ESTs encoded a protein that shared sequence similarity with a venom protein of unknown function potentially specific of the <it>Chelonus </it>lineage. Venom components specific to <it>C. inanitus </it>included a C-type lectin domain containing protein, a chemosensory protein-like protein, a protein related to yellow-e3 and ten new proteins which shared no significant sequence similarity with known sequences. In addition, several venom proteins potentially able to interact with chitin were also identified including a chitinase, an imaginal disc growth factor-like protein and two putative mucin-like peritrophins.</p> <p>Conclusions</p> <p>The use of the combined approaches has allowed to discriminate between cellular and truly venom proteins. The venom of <it>C. inanitus </it>appears as a mixture of conserved venom components and of potentially lineage-specific proteins. These new molecular data enrich our knowledge on parasitoid venoms and more generally, might contribute to a better understanding of the evolution and functional diversity of venom proteins within Hymenoptera.</p

Social Transfer of Pathogenic Fungus Promotes Active Immunisation in Ant Colonies

Social contact with fungus-exposed ants leads to pathogen transfer to healthy nest-mates, causing low-level infections. These micro-infections promote pathogen-specific immune gene expression and protective immunization of nest-mates

Role and evolution of virus genes involved in a host-parasitoid interaction

Les symbioses, en permettant l’acquisition de nouvelles fonctions, ont joué un rôle majeur dans l’évolution des organismes. Cette thèse, vise à comprendre les mécanismes moléculaires et évolutifs qui font des polydnavirus des symbiontes indispensables au succès parasitaire de guêpes parasitoïdes. Pour cela, nous avons étudié l’évolution des gènes viraux et les fonctions physiologiques potentiellement ciblées chez l’hôte. Nous avons montré que l’évolution des gènes viraux était caractérisée par des duplications et une divergence rapide, et répondait à des pressions de sélection positive. Chez l’hôte, nous avons montré que des protéases à cystéine, potentiellement ciblées par des gènes viraux, étaient régulées au cours du parasitisme au niveau de la transcription et de la traduction, suggérant que ces protéines doivent jouer un rôle important au cours du parasitisme. Il reste maintenant à déterminer quelle est la fonction de ces protéines et leur influence sur l’évolution des gènes viraux.Symbioses have largely contributed to the evolution of species by providing new functions enabling niche colonization. Here, our aim was to study the molecular evolution of virulence factors encoded by polydnaviruses engaged in a mutualist association with parasitoid wasps and which are essential for parasitism success. We studied both PDV genes and their Lepidopteran host targets to determine how these genes evolved and the host functions targeted. We showed that natural selection has largely contributed to the evolution of cystatin and protein-tyrosine- phosphatase viral genes. 3D structure modelling showed that selection acted in cystatin active sites. Moreover, we underlined the dynamic evolution of PDV genes mainly explained by gene duplication processes. In the host, potential cystatin targets, cysteine proteases, were shown to be regulated during parasitism both at the gene and the protein level. These results emphasized the important role played by these proteins against invaders. To understand both evolutionary and mechanistic processes involved in this interaction, it is now necessary to determine the function of cysteine proteases and to study how virus gene evolution could be shaped by host defence factors

Rôle et évolution de facteurs de virulence impliqués dans une interaction hôte-parasitoïde

Symbioses have largely contributed to the evolution of species by providing new function. Here, our aim was to study the molecular evolution of virulence factors of polydnaviruses, essential symbiont for wasp parasitism success. We studied both PDV genes and their Lepidopteran host targets to determine how these genes evolved and the host functions targeted. We showed that natural selection has largely contributed to virus genes evolution. We also underlined the dynamic evolution of PDV genes mainly explained by gene duplication processes. In the host, cysteine proteases, were shown to be regulated during parasitism both at the gene and the protein level. These results emphasized the important role played by these proteins against invaders. To understand both evolutionary and mechanistic processes involved in this interaction, it is now necessary to determine the functionLes associations mutualistes, en permettant l'acquisition de nouvelles fonctions, ont joué un rôle majeur dans l'évolution des espèces. Pour comprendre en quoi ces associations sont impliquées dans l'adaptation des espèces nous avons étudié un cas unique de mutualisme associant un virus de type polydnavirus et une guêpe parasitoïde. Dans cette association, le virus est injecté dans l'hôte lépidoptère lors de l'oviposition et joue un rôle majeur dans le succès parasitaire en induisant une altération des fonctions physiologiques de l'hôte. En regard du nombre d'espèces de guêpes caractérisées par cette association, le virus doit constituer une innovation adaptative majeure et jouer un rôle déterminant dans l'évolution et la diversification des espèces de guêpes. En quoi cette association joue t-elle un rôle déterminant dans l'évolution et l'adaptation des guêpes ? Quelles sont les fonctions physiologiques ciblées chez l'hôte ? Pour répondre à ces questions nous avons étudié l'évolution de deux familles de gènes codant pour des facteurs de virulence potentiels et nous avons exploré les fonctions physiologiques d'une protéine potentiellement ciblée lors du parasitisme. Nous avons mis en évidence le rôle important de la sélection naturelle dans l'évolution des familles de gènes viraux. Par modélisation de la structure tridimensionnelle d'un facteur de virulence codant pour des cystatines, nous avons montré que cette sélection agissait préférentiellement au niveau des sites d'interaction avec les protéines cibles. De plus, cette étude souligne le caractère dynamique de l'évolution des facteurs de virulence incluant de multiples évènements de duplication, caractérisés par des processus de perte et d'acquisition au cours de l'évolution de l'association. Le caractère adaptatif et dynamique de l'évolution des gènes viraux a aussi été étudié en regard de l'évolution des espèces de guêpes et de leur spectre d'hôte. Par une approche fonctionnelle, nous avons étudié le rôle physiologique de protéases à cystéine qui constituent des cibles potentielles des cystatines virales. Nous avons montré que ces protéases sont régulées spécifiquement au cours du parasitisme au niveau protéique et transcriptionnel. Nous avons également montré que l'activité de ces protéases est modifiée après parasitisme. L'évolution adaptative et dynamique des facteurs de virulence reflètent leur rôle important dans le parasitisme. Il reste maintenant à montrer comment ces facteurs interagissent sur la physiologie de l'hôte lépidoptère. Des protéases à cystéine sont spécifiquement ciblées par le parasitisme, en étudiant les mécanismes d'interaction de ces protéases avec les cystatines virales et les processus coévolutifs mis en jeu, nou

Evolution and Origin of Polydnavirus Virulence Genes. In Symbionts and Pathogens

International audiencePolydnaviruses (PDVs) have a unique life-cycle, comprised of a mutualistic lifestyle with their associated parasitoid wasps and a parasitic interaction with the lepidopteran wasp hosts. They are present as proviruses in the parasitoid wasps that harbor them. The female wasps produce particles that contain circular double-stranded DNA versions of the viruses that are injected into the wasps' lepidopteran hosts. This 'injected circular form' is replication deficient but is absolutely essential for the physiological regulation of caterpillars that leads to parasitoid survival. PDVs are divided into two genera, bracoviruses (BVs) and ichnoviruses (IVs) associated with tens of thousands of endoparasitoid braconid and ichneumonid wasps, respectively, belonging to the Ichneumonoidea superfamily. The absence of PDVs in basal lineages of Ichneumonoidea strongly suggests that the association of BVs with braconids and IVs with ichneumonids arose independently. In ichneumonids, PDVs have been identified in the Campopleginae and Banchinae subfamily. The unique genomic features of the first banchine virus examined to date also suggest they could have an origin distinct from those of IVs and BVs (Lapointe et al., 2007). The absence of genes involved in virus production in the injected circular form is likely to be a signature of the reductive evolution that these PDVs have been subjected to, due to their reliance on wasps for transmission and the absence of replication in host larvae. In this chapter, we will focus on the PDV circular genomes that are injected into the lepidopteran hosts and highlight how selection pressures are likely to have modeled their content, organization, gene function, and efficiency. These PDV genomes are atypical for viruses in the sense that they harbor numerous genes, many of whic

Transfer of a chromosomal Maverick to endogenous bracovirus in a parasitoid wasp

International audienceBracoviruses are used by parasitoid wasps to allow development of their progeny within the body of lepidopteran hosts. In parasitoid wasps, the bracovirus exists as a provirus, integrated in a wasp chromosome. Viral replication occurs in wasp ovaries and leads to formation of particles containing dsDNA circles (segments) that are injected into the host body during wasp oviposition. We identified a large DNA transposon Maverick in a parasitoid wasp bracovirus. Closely related elements are present in parasitoid wasp genomes indicating that the element in CcBV corresponds to the insertion of an endogenous wasp Maverick in CcBV provirus. The presence of the Maverick in a bracovirus genome suggests the possibility of transposon transfers from parasitoids to lepidoptera via bracoviruses

Speciation in Thaparocleidus (Monogenea : Dactylogyridae) parasitizing Asian pangasiid catfishes

The phylogeny of monogeneans of the genus Thaparocleidus that parasitize the gills of Pangasiidae in Borneo and Sumatra was inferred from molecular data to investigate parasite speciation. The phylogeny of the Pangasiidae was also reconstructed in order to investigate host-parasite coevolutionary history. The monophyly of Thaparocleidus parasitizing Pangasiidae was confirmed. Low intraspecies molecular variability was observed in three Thaparocleidus species collected from geographically distant localities. However, a high intraspecies molecular variability was observed in two Thaparocleidus species suggesting that these species represent a complex of species highly similar in morphology. Distance-based and tree-based methods revealed a significant global fit between parasite and host phylogenies. Parasite duplication (i.e., intrahost speciation) was recognized as the most common event in Thaparocleidus, while the numbers of cospeciation and host switches were lower and similar to each other. When collapsing nodes correspond to duplication cases, our results suggest host switches in the Thaparocleidus-Pangasiidae system precluding congruence between host and parasite trees. We found that the morphometric variability of the parasite attachment organ is not linked to phylogeny, suggesting that the attachment organ is under adaptive constraint. We showed that haptor morphometry is linked to host specificity, whereby nonspecific parasites display higher morphometric variability than specialists

Evolutionary mechanisms driving the evolution of a large polydnavirus gene family coding for protein tyrosine phosphatases

Background: Gene duplications have been proposed to be the main mechanism involved in genome evolution and in acquisition of new functions. Polydnaviruses (PDVs), symbiotic viruses associated with parasitoid wasps, are ideal model systems to study mechanisms of gene duplications given that PDV genomes consist of virulence genes organized into multigene families. In these systems the viral genome is integrated in a wasp chromosome as a provirus and virus particles containing circular double-stranded DNA are injected into the parasitoids' hosts and are essential for parasitism success. The viral virulence factors, organized in gene families, are required collectively to induce host immune suppression and developmental arrest. The gene family which encodes protein tyrosine phosphatases (PTPs) has undergone spectacular expansion in several PDV genomes with up to 42 genes. Results: Here, we present strong indications that PTP gene family expansion occurred via classical mechanisms: by duplication of large segments of the chromosomally integrated form of the virus sequences (segmental duplication), by tandem duplications within this form and by dispersed duplications. We also propose a novel duplication mechanism specific to PDVs that involves viral circle reintegration into the wasp genome. The PTP copies produced were shown to undergo conservative evolution along with episodes of adaptive evolution. In particular recently produced copies have undergone positive selection in sites most likely involved in defining substrate selectivity. Conclusion: The results provide evidence about the dynamic nature of polydnavirus proviral genomes. Classical and PDV-specific duplication mechanisms have been involved in the production of new gene copies. Selection pressures associated with antagonistic interactions with parasitized hosts have shaped these genes used to manipulate lepidopteran physiology with evidence for positive selection involved in adaptation to host targets