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Hajdusek, O.

Publications and source records attributed to Hajdusek, O..

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Host-derived lipids license proliferation of sterol-auxotrophic tick cells and promote flavivirus output

Sterol molecules play indispensable roles in the biology of cells and organisms. Yet, arthropods, including ticks, lack the capacity for de novo sterol biosynthesis and must rely on exogenous sources. How these organisms sense and utilise exogenous lipids at the cellular level remains poorly understood. Here, we show that host cholesterol and linoleic acid act as essential external signals that license proliferation of Ixodes ricinus tick cells. In lipid-depleted conditions, cells fail to proliferate despite retaining intracellular cholesterol, indicating that exogenous lipids function as regulatory cues rather than merely structural components. We further demonstrate that tick cells exhibit a non-canonical intracellular distribution of cholesterol, with predominant accumulation in lysosome-associated compartments, and that lipid availability induces a coordinated transcriptional program involving lipid uptake and trafficking machinery. Importantly, we find that limiting host lipid availability significantly reduces extracellular RNA output of tick-borne encephalitis virus (TBEV) from infected tick cells, indicating that viral production is affected by the metabolic state of the host cell. Together, our findings establish host-derived lipids as key regulators of cellular state in a sterol-auxotrophic organism and identify lipid availability as a determinant of flavivirus output. This work provides a conceptual framework linking nutrient sensing, cell proliferation, and vector-pathogen interactions.

microbiology↗

The Bacterial Kinase AnmK Integrates into Tick Genome and Biology

Horizontal Gene Transfer (HGT) contributes to eukaryotic evolution, potentially bringing phenotypic novelties to the recipient organisms. Ticks pose a severe threat to human health as vectors of various pathogens, including viruses, bacteria and protozoa, while stably hosting endosymbiotic bacteria. As such, these obligate blood-feeding parasites have been and continue to be exposed to HGT from a broad range of donors. To determine whether bacterial-to-tick HGT has contributed to important tick traits, we surveyed Ixodes tick genomes for HGT events. We revealed duplications of the known bacteria-derived gene dae2 and discovered two novel cases of bacterial HGT, the most remarkable of which involves a bacterial peptidoglycan metabolic gene (anmK) acquired by the common ancestor of ticks. The acquisition of an intron demonstrates "eukaryotization" of anmK within tick genomes. Transcript profiling revealed that anmK expression is upregulated during blood feeding, peaking in female ovaries, a niche occupied by horizontally acquired endosymbionts. Biochemical analysis confirmed that, to some extent, recombinant AnmK retains kinase activity on its cognate substrate - the bacterial cell wall component 1,6-anhydro-N-acetylmuramic acid. Immunolocalization showed that the enzyme is predominantly localized towards outer layers of the vitellogenic oocytes. Silencing of anmK in different tick species compromised blood-feeding and reproduction, demonstrating that this domesticated bacterial enzyme underpins reproductive fitness across tick species. Our findings exemplify the ability of horizontally acquired genes to integrate into the host biology and thereby shape host life history.

evolutionary biology↗

A newly identified three-domain C-type lectin associated with blood feeding in the tick Ixodes ricinus

Ixodes ricinus ticks are widely distributed throughout Europe and represent major vectors of tick-borne encephalitis virus and the Lyme borreliosis agent Borrelia burgdorferi sensu lato. In invertebrates, C-type lectins are commonly associated with innate immune functions, and several such lectins have been predicted in I. ricinus. Given the limited knowledge of lectin function in ticks, we characterized three carbohydrate-recognition domains (CRDs) of a novel C-type lectin identified in the I. ricinus transcriptome (IrCLec). The tertiary structures of CRD1, CRD2, and CRD3, predicted using the AlphaFold 3 program, corresponded to the typical structure of C-type lectins. Conserved carbohydrate-binding motifs were identified in CRD3, whereas non-canonical motifs were present in CRD1 and CRD2. Recombinant His-tagged CRDs were produced and analysed for carbohydrate-binding activity. Glycan array analysis revealed binding of all three domains to selected glycans, while hemagglutination assays demonstrated pronounced binding activity of CRD1 and CRD2 toward human erythrocyte antigens of blood groups A, B, and O. IrCLec expression was highest in the tick midgut and also detected in hemocytes, with expression levels increasing after blood feeding. RNAi-mediated silencing of IrCLec impaired blood feeding efficiency in tick nymphs. Together, these results indicate that IrCLec plays an important role in blood feeding and may additionally participate in lectin-mediated host-pathogen or host-blood component interactions.

biochemistry↗

Integrative assessment of the transcriptome and virome of the poultry red mite Dermanyssus gallinae.

Dermanyssus gallinae is a blood-feeding mite that parasitises on wild birds and farmed poultry. The D. gallinae mite has a short life cycle of fewer than two weeks from the egg to an egg-laying female. The remarkably swift processing of blood, together with the capacity to blood-feed in most developmental stages, makes this mite a highly debilitating pest. We have constructed developmental stage-specific transcriptomes, through Illumina RNA-seq, to mine the repertoire of protein-encoding mRNA transcripts, products of which participate in key processes that ensure the success of blood digestion, rapid ontogeny, and immunity. As a result of high reproductive capacity, the prevalence of D. gallinae in egg-producing poultry farms globally causes significant economic losses. Acaricides that are used to limit the reproduction of D. gallinae mites target cys-loop ion channels, which are widely shared across the phylogeny of invertebrates. To catalogue a comprehensive list of potential invertebrate-specific ion channels, we have constructed and analysed an additional RNA-seq library of D. gallinae micro-dissected midguts, a tissue with direct exposure to host blood and potential anti-parasitics. We phylogenetically defined groups of cys-loop proteins and probed their sensitivity to selected acaricides. Ultimately, we have catalogued all assembled transcripts and their expression values in a hyper-linked excel sheet with available sequences of individual contigs. The transcriptomic data were complemented by mass-spectrometry (MS)-based metabolite identification and by viability assays using selected inhibitors applied either by microinjection or through artificial membrane feeding. Additionally, we have described the RNA-virome of D. gallinae and identified a novel virus dubbed Red Mite Quaranjavirus 1.

microbiology↗

Tracking Borrelia afzelii from infected Ixodes ricinus nymphs to mice suggests a direct ‘gut-to-mouth’ route of Lyme disease transmission

Quantitative tracking of Borrelia afzelii has shown that its transmission cycle differs from the salivary route of B. burgdorferi transmission by Ixodes scapularis. Borrelia afzelii are abundant in the guts of unfed Ixodes ricinus nymphs and their numbers continuously decrease during feeding. In contrast, spirochetes are not present in the salivary glands. Borrelia afzelii transmission starts during the early stages of feeding, spirochetes could be detected in murine skin within 1 day of tick attachment. Tick saliva is not essential for B. afzelii infectivity, the main requirement for successful host colonization being a change in outer surface protein expression that occurs in the tick gut during feeding. Spirochetes in vertebrate mode are able to survive within the host even if the tick is not present. On the basis of our data we propose that a direct gut-to-mouth route of infection appears to be the main route of B. afzelii transmission.\n\nImportanceLyme borreliosis is the most common vector-borne disease in the USA and Europe. The disease is caused by the Borrelia spirochetes and is transmitted through Ixodes ticks. A better understanding of how Borrelia spirochetes are transmitted is crucial for development of efficient vaccines for preventing Lyme borreliosis. Here we present that the transmission of European B. afzelii spirochetes by I. ricinus ticks significantly differs from the model transmission cycle described for American B. burgdorferi/I. scapularis. We suggest that B. afzelii is not transmitted via salivary glands but most likely through the midgut to mouthpart route. We further demonstrate that tick saliva is not important for B. afzelii transmission and infectivity. Therefore, we support early studies by Willy Burgdorfer, who proposed that Borrelia transmission occurs by regurgitation of infected gut contents. Our findings collectively point to the Borrelia-tick midgut interface as the correct target in our endeavours to combat Lyme borreliosis.

microbiology↗