Duplais, C. | bioRxiv Science

bioRxiv2020

Heterodimeric insecticidal peptide provides new insights into the molecular and functional diversity of ant venoms

Ants use venom for predation, defence and communication, however, the molecular diversity, function and potential applications of ant venom remains understudied compared to other venomous lineages such as arachnids, snakes and cone snails. In this work, we used a multidisciplinary approach that encompassed field work, proteomics, sequencing, chemical synthesis, structural analysis, molecular modelling, stability studies, and a series of in vitro and in vivo bioassays to investigate the molecular diversity of the venom of the Amazonian Pseudomyrmex penetrator ants. We isolated a potent insecticidal heterodimeric peptide {Delta}-pseudomyrmecitoxin-Pp1a ({Delta}-PSDTX-Pp1a) composed of a 27-residue long A-chain and a 33-residue long B-chain crosslinked by two disulfide bonds in an antiparallel orientation. We chemically synthesised {Delta}-PSDTX-Pp1a, its corresponding parallel AA and BB homodimers, and its monomeric chains and demonstrated that {Delta}-PSDTX-Pp1a had the most potent insecticidal effects in blow fly assays (LD50 = 3 nM). Molecular modelling and circular dichroism studies revealed strong alpha-helical features, indicating its cytotoxic effects could derive from membrane disruption, which was further supported by insect cell calcium assays. The native heterodimer was also substantially more stable against proteolytic degradation (t1/2 =13 h) than its homodimers or monomers (t1/2 <20 min), indicating an evolutionary advantage of the more complex structure. The proteomic analysis of Pseudomyrmex penetrator venom and in-depth characterisation of {Delta}-PSDTX-Pp1a provide novel insights in the structural complexity of ant venom, and further exemplifies how nature exploits disulfide-bond formation and dimerization to gain an evolutionary advantage via improved stability; a concept that is also highly relevant for the design and development of peptide therapeutics, molecular probes and bioinsecticides.

pharmacology and toxicology↗

bioRxiv2020

Assessing biosynthetic gene cluster diversity in a multipartite nutritional symbiosis between herbivorous turtle ants and conserved gut symbionts

In insect-microbe nutritional symbioses the symbiont supplements the low nutrient diet of the host by producing amino acids and vitamins, and degrading lignin or polysaccharides. In multipartite mutualisms composed of multiple symbionts from different taxonomical orders, it has been suggested that in addition to the genes involved in the nutritional symbiosis the symbionts maintain genes responsible for the production of metabolites putatively playing a role in the maintenance and interaction of the bacterial communities living in close proximity. To test this hypothesis we investigated the diversity of biosynthetic gene clusters (BGCs) in the genomes and metagenomes of obligate gut symbionts associated with the herbivorous turtle ants (genus: Cephalotes). We studied 17 Cephalotes species collected across several geographical areas to reveal that (i) mining bacterial metagenomes and genomes provides complementary results demonstrating the robustness of this approach with metagenomic data, (ii) symbiotic gut bacteria have a high diversity of BGCs which is correlated with host geography but not host phylogeny, (iii) the majority of the BGCs comes from the bacteria involved in the nutritional symbiosis supporting conserved metabolic functions for colonization, communication and competition in the gut environment, (iv) phylogenetic analysis of arylpolyene, polyketide (PK), and siderophore shows high similarity between BGCs of a single symbiont across different ant host species, while non-ribosomal peptide (NRP) shows high similarity between BGCs from different bacterial orders within a single host species suggesting multiple mechanisms for genome evolution of these obligate mutualistic gut bacteria.

evolutionary biology↗