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Biology subjects

Luksan, O.

Publications and source records attributed to Luksan, O..

3 recordsLinked to original sources

Identification of the trail-following pheromone receptor in termites

Chemical communication is the cornerstone of eusocial insect societies since it mediates the social hierarchy, division of labor, and concerted activities of colony members. The chemistry of social insect pheromones received considerable attention in both major groups of social insects, the eusocial Hymenoptera and termites. By contrast, current knowledge on molecular mechanisms of social insect pheromone detection by odorant receptors (ORs) is limited to hymenopteran social insects and no OR was yet functionally characterized in termites, the oldest eusocial insect clade. Here, we present the first OR deorphanization in termites. We selected four OR sequences from the previously annotated antennal transcriptome of the termite Prorhinotermes simplex (Rhinotermitidae), expressed them in Empty Neuron Drosophila, and functionally characterized using single sensillum recording (SSR) and a panel of termite semiochemicals. In one of the selected ORs, PsimOR14, we succeeded in obtaining strong and reliable responses to the main component of P. simplex trail-following pheromone, the monocyclic diterpene neocembrene. PsimOR14 showed a narrow tuning to neocembrene; only one additional compound out of 67 tested (geranylgeraniol) generated non-negligible responses. Subsequently, we used SSR in P. simplex workers and identified the olfactory sensillum specifically responding to neocembrene, thus likely expressing PsimOR14. We report on homology-based modelling of neocembrene binding by PsimOR14 and show how different ligands impact the receptor dynamicity using molecular dynamics simulations. Finally, we demonstrate that PsimOR14 is significantly more expressed in worker antennae compared to soldiers, which correlates with higher sensitivity of workers to neocembrene.

animal behavior and cognition↗

Extended longevity of termite kings and queens is accompanied by extranuclear localization of telomerase in somatic organs and caste-specific expression of its isoforms

Kings and queens of termites are endowed with an extraordinary longevity coupled with lifelong fecundity. We recently reported that termite kings and queens display a dramatically increased enzymatic activity and abundance of telomerase in their somatic organs when compared to short-lived workers and soldiers. We hypothesized that this telomerase activation may represent a non-canonical pro-longevity function, independent of its canonical role in telomere maintenance. Here, we explore this avenue and investigate whether the presumed non-canonical role of telomerase may be due to alternative splicing of the catalytic telomerase subunit TERT and whether the subcellular localization of TERT isoforms differs among organs and castes in the termite Prorhinotermes simplex. We empirically confirm the expression of four in silico predicted splice variants (psTERT1-A, psTERT1-B, psTERT2-A, psTERT2-B), defined by N-terminal splicing implicating differential localizations, and C-terminal splicing giving rise to full-length and truncated isoforms. We show that the transcript proportions of the psTERT are caste- and tissue-specific and that the extranuclear full-length isoform TERT1-A is relatively enriched in the soma of neotenic kings and queens compared to their gonads and to the soma of workers. We also show that extranuclear TERT protein quantities are significantly higher in the soma of kings and queens compared to workers, namely due to the cytosolic TERT. Independently, we confirm by microscopy the extranuclear TERT localization in somatic organs. We conclude that the presumed pleiotropic action of telomerase combining the canonical nuclear role in telomere maintenance with extranuclear functions is driven by complex TERT splicing.

molecular biology↗

Exploring RNA modifications in infectious non-coding circular RNAs

Viroids, small circular non-coding RNAs, act as infectious pathogens in higher plants, demonstrating high stability despite consisting solely of naked RNA. Their dependence of replication on host machinery poses the question of whether RNA modifications play a role in viroid biology. Here, we explore RNA modifications in the avocado sunblotch viroid (ASBVd) and the citrus exocortis viroid (CEVd), representative members of viroids replicating in chloroplasts and the nucleus, respectively, using LC-MS and Oxford Nanopore Technology (ONT) direct RNA sequencing. Although no modification was detected in ASBVd, CEVd contained approximately one m6 A per RNA molecule. ONT sequencing predicted three m6 A positions. Employing orthogonal SELECT method, we confirmed m6 A in two positions A353 and A360, which are highly conserved among CEVd variants. These postitions are located in the left terminal region of the CEVd rod-like structure where likely RNA Pol II and and TFIIIA-7ZF bind, thus suggesting potential biological role of methylation in viroid replication. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=125 SRC="FIGDIR/small/584625v2_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@1dfe2f1org.highwire.dtl.DTLVardef@1ac717eorg.highwire.dtl.DTLVardef@4f1563org.highwire.dtl.DTLVardef@542d9_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗