Biology subjects

Fuchs, R.

Publications and source records attributed to Fuchs, R..

9 recordsLinked to original sources

bioRxiv2026

KLHL9 acts as a KSHV vBcl-2-interacting host factor that supports lytic replication

Kaposis sarcoma-associated herpesvirus (KSHV; human herpesvirus 8) is an oncogenic gammaherpesvirus that causes Kaposis sarcoma and several lymphoproliferative disorders. The KSHV open reading frame (ORF) 16 encodes viral Bcl-2 (vBcl-2), which inhibits apoptosis and autophagy. vBcl-2 is required for efficient lytic replication and production of infectious progeny; however, this essential role is independent of its established functions regulating cell death. To identify host factors that may contribute to vBcl-2 function, we employed proteomic analysis of HA-vBcl-2 immunoprecipitates from lytically reactivated KSHV-infected cells. This analysis identified the BTB-Kelch protein KLHL9, a substrate-specific adaptor for Cullin 3-RING ligase (CRL3) complex, as a candidate vBcl-2-associated factor. We validated the vBcl-2-KLHL9 association by reciprocal co-immunoprecipitation from infected cells, and by ectopic expression in uninfected cells. During lytic reactivation, KLHL9 redistributed from a predominantly cytoplasmic/perinuclear pattern to a mitochondrial pattern that overlapped with HA-vBcl-2. Alanine-scanning mutagenesis mapped a KLHL9-interaction determinant to the N-terminal region of vBcl-2, and vBcl-2 mutants defective in binding failed to colocalize with KLHL9. Conversely, KLHL9 deletion analysis implicated the C-terminal Kelch-repeat region in efficient association with vBcl-2. AlphaFold modeling supported the interface regions. However, vBcl-2 did not show detectable KLHL9-dependent ubiquitination, suggesting that it is unlikely to be a substrate of a KLHL9-containing complex. Functionally, CRISPR/Cas9-mediated KLHL9 disruption reduced lytic viral protein accumulation and infectious progeny production, while re-expression of sgRNA-resistant KLHL9 partially restored these phenotypes. Together, these findings suggest that vBcl-2 may engage host CRL3 complexes during productive infection, and identify KLHL9 as a vBcl-2-associated host factor that supports efficient KSHV lytic replication. Author SummaryKaposis sarcoma-associated herpesvirus (KSHV) is a cancer-associated virus that alternates between a latent cycle and a productive lytic cycle, during which new infectious virus particles are made. The viral protein vBcl-2 is best known for regulating cell-death and autophagy pathways, but earlier studies showed that it is required for efficient virus production through additional, non-canonical functions. In this study, we identify the cellular protein KLHL9 as a host factor that associates with KSHV vBcl-2. KLHL9 normally functions as a specific adaptor in protein ubiquitination pathways that regulate the turnover, localization, or activity of target proteins through ubiquitination. We show that vBcl-2 and KLHL9 associate with one another, and that KLHL9 redistributes toward the mitochondria in cells expressing vBcl-2. Importantly, loss of KLHL9 impairs the expression of viral lytic proteins and reduces the production of new infectious viruses. This defect resembles the phenotype caused by loss of vBcl-2, suggesting that KSHV employs vBcl-2 to engage KLHL9-dependent host machinery during productive infection. Understanding this interaction may help reveal how herpesviruses reprogram infected cells to create an environment that supports virus production.

microbiology↗

bioRxiv2025

Metabolic biomarker-based phenotyping unveils quantitative effects of plant resistance and pathogen aggressiveness in the grapevine (Vitis spp.) - downy mildew (Plasmopara viticola) pathosystem.

Grapevine resistance to downy mildew has been primarily associated with major "Resistance to Plasmopara viticola" (Rpv) loci, which are extensively used in breeding programs. Resistant varieties represent an effective solution to mitigate the environmental impact of fungicide application in viticulture, but P. viticola strains able to overcome major Rpv have become a main threat to their cultivation. Pyramiding resistance loci in the same variety enhances plant resistance, but interactions involving stacked and defeated Rpv and different P. viticola strains are poorly documented. Investigation of these interactions may uncover new information for the development of efficient breeding strategies, the optimal exploitation of Rpv, and the building of durable resistance. In the present study, a grapevine offspring carrying single and pyramided Rpv1, Rpv3.1 and Rpv10 was phenotyped in laboratory conditions for the resistance to P. viticola using a naive strain and a strain virulent towards Rpv10. By using a high-resolution phenotyping strategy based on P. viticola metabolic biomarkers, we demonstrated that the efficacy of Rpv combinations and aggressiveness of P. viticola strains can be quantified in the early phase of infection and were often related to sporulation outcome. Furthermore, we described how a limited residual effect of a defeated Rpv may become significant in pyramiding. In conclusion, in addition to providing the keys to streamlining resistance utilization in grapevine, our research presents a distinctive case study that provides valuables information for breeding new resistant varieties, thanks to an innovative "omic"-based phenotyping approach, which may be adapted to other plant pathosystems.

plant biology↗

bioRxiv2025

Coronavirus protein interaction mapping in bat and human cells identifies molecular and genetic switches for immune evasion and replication

Coronaviruses, including SARS-CoV-2, can cause severe disease in humans, whereas reservoir hosts like Rhinolophus bats remain asymptomatic. To investigate how host-specific protein-protein interactions (PPIs) influence infection, we generated comparative PPI maps for SARS-CoV-2 and its bat-origin relative RaTG13 using affinity purification-mass spectrometry (AP-MS) in human and Rhinolophus ferrumequinum (RFe) bat cells. This approach identified both conserved and virus- and host-specific interactions that regulate infection dynamics. Notably, SARS-CoV-2 required a non-synonymous mutation in nucleocapsid to replicate in bat cells expressing human ACE2 and TMPRSS2. Analysis of the viral protein Orf9b revealed differential interactions with mitochondrial proteins Tom70 and MTARC2. A single residue difference in Orf9b between SARS-CoV-2 and RaTG13 functions as a molecular switch, weakening Tom70 binding and immune evasion in human cells while enhancing interaction with the bat-specific restriction factor MTARC2. These findings demonstrate how a single-residue substitution can reshape virus-host interactions and contribute to immune evasion and host adaptation.

systems biology↗

bioRxiv2025

Breeding for durable resistance in crops: defeated loci may act as Trojan horses compromising the effectiveness of major resistance genes.

Resistance breeding offers invaluable perspectives for environment-friendly crop protection, but its success may be limited by the breakdown of plant resistance by pathogen strains. This threat is particularly acute for perennial crops, which may be cultivated for several decades. With the increasing use of new varieties carrying multiple major resistance loci, grapevine (Vitis spp.) represents a distinctive model to investigate the broad agreement that combining several resistance genes (pyramiding) enhances both resistance efficacy and durability. To this end, grapevine progenies segregating for four resistance loci against Plasmopara viticola (Rpvs) were used to evaluate the efficiency of single and pyramided major loci when confronted to naive and Rpv-breaking pathogen strains. In the context of polygenic resistance, both undefeated and defeated Rpvs provided significant quantitative effects. However, interactions between pyramided Rpvs were either beneficial, neutral or detrimental to the level of resistance, depending on the loci combination and pathogen strain. In particular, the fact that the presence of defeated resistance loci may compromise the resistance provided by functional major loci has important implications for crops resistance breeding. Thorough phenotypic investigations of pyramiding breeding schemes emerge as a critical step for the effective and durable management of genetic resistances and plant diseases.

plant biology↗

bioRxiv2024

Investigating the impact of edge weight selection on the pig trade network topology

Traceability of animal movements and robust surveillance are crucial for identifying and controlling animal diseases. Risk-based surveillance, i.e. network-based approaches, can identify higher-risk holdings or trades. However, node ranking, useful to identify "influential" nodes (holdings) in the network, varies with the considered metrics. We use a dataset of pig movements in Upper Austria from 2021 to study the robustness of node ranking through three centrality metrics and compare them with epidemic model ranking. Incorporating edge weights may influence the network analysis, therefore, we simulate two representations using edge weights based on: i) the frequency of exchanges between holdings ("frequency-based") and ii) the number of pigs exchanged ("volume-based"). We compare the impact of the edge weight on the network topology, community structure, and node ranking in a network with 5,766 nodes and 92,914 edges. Results revealed distinct edge weight distributions: frequency-based network exhibited a bimodal pattern, while volume-based was more uniform. Strength centrality exhibited the highest correlation with simulation-based rankings, particularly for the top 5% highest-ranked nodes ({tau}b = 0.51 for frequency-based and{tau} b = 0.5 for volume-based). These findings highlight that using strength centrality to identify critical nodes can significantly enhance surveillance strategies, making them more efficient and field-deployable, enhancing traditional methods without requiring extensive simulations. Author summaryEarly detection of infectious diseases through surveillance activities is important to prevent severe impacts on the livestock sector and avoid major economic losses that often result from delayed detection. Prioritizing surveillance efforts through a data-driven approach presents a strategic advantage compared to random sampling. Here, we propose two network representations of the Upper Austrian pig trade network 2021, which show livestock holdings as nodes and animal movements as links between these nodes, which can represent the volume of pigs exchanged or the frequency of these exchanges. Using network analysis methods, we identify "influential" holdings in the network, i.e., those playing a key role in the network, either due to their numerous connections with other holdings, their position on many trade paths, or their crucial role in disease transmission dynamics. We show that using strength centrality can effectively identify key holdings for targeted surveillance. Adopting network-based surveillance can facilitate resource allocation for veterinary surveillance programs, offering a cost-effective strategy for disease management, enabling tailored, more effective, and timely interventions.

bioinformatics↗

bioRxiv2023

A multi-omics strategy for the study of microbial metabolism: application to the human skin's microbiome

Although metabolomics data acquisition and analysis technologies have become increasingly sophisticated over the past 5-10 years, deciphering a metabolites function from a description of its structure and its abundance in a given experimental setting is still a major scientific and intellectual challenge. To point out ways to address this data to knowledge challenge, we developed a functional metabolomics strategy that combines state-of-the-art data analysis tools and applied it to a human scalp metabolomics dataset: Skin swabs from healthy volunteers with normal or oily scalp (Sebumeter score 60-120, n = 33; Sebumeter score > 120, n = 41) were analyzed by LC-MS/MS, yielding four metabolomics datasets for RP18 or HILIC separation in ESI+ or ESI- ionization mode. Following our data analysis strategy, we were able to obtain increasingly comprehensive structural and functional annotations, by applying the GNPS (Wang et al., Nat. Biotechnol. 2016), SIRIUS (Duhrkop et al. Nat. Methods 2019), and MicrobeMASST (Zuffa et al., bioRxiv 2023) tools. We finally combined the metabolomics data with a corresponding metagenomic sequencing dataset using MMvec (Morton et al., Nat. Methods 2019), gaining insights into the metabolic niche of one of the most prominent microbes on the human skin, Staphylococcus epidermidis. IMPORTANCESystems biology research on host-associated microbiota focuses on two fundamental questions: Which microbes are present and how do they interact with each other, their host, and the broader host environment? Metagenomics provides us with a direct answer to the first part of the question: it unveils the microbial inhabitants, e.g., on our skin, and can provide insight into their functional potential. Yet, it falls short in revealing their active role. Metabolomics shows us the chemical composition of the environment in which microbes thrive and the transformation products they produce. In particular, untargeted metabolomics has the potential to observe a diverse set of metabolites and is thus an ideal complement to metagenomics. However, this potential often remains underexplored due to the low annotation rates in MS-based metabolomics and the necessity for multiple experimental chromatographic and mass spectrometric conditions. Beyond detection, prospecting metabolites functional role in the host/microbiome metabolome requires identifying the biological processes and entities involved in their production and biotransformations. In the present study of the human scalp, we developed a strategy to achieve comprehensive structural and functional annotation of the metabolites in the human scalp environment, thus diving one step deeper into the interpretation of omics data. Leveraging a collection of openly accessible software tools and integrating microbiome data as a source of functional metabolite annotations, we finally identified the specific metabolic niche of Staphylococcus epidermidis, one of the key players of the human skin microbiome.

systems biology↗

bioRxiv2021

How do symbiotic associations in lecideoid lichens respond to different environmental conditions along the Transantarctic Mountains, Ross Sea region, Antarctica?

Lecideoid lichens as dominant vegetation-forming organisms in the climatically harsh areas of the southern part of continental Antarctica show clear preferences in relation to environmental conditions (i.e. macroclimate). 306 lichen samples were included in the study, collected along the Ross Sea coast (78{degrees}S-85.5{degrees}S) at six climatically different sites. The species compositions as well as the associations of their two dominant symbiotic partners (myco- and photobiont) were set in context with environmental conditions along the latitudinal gradient. Diversity values were nonlinear with respect to latitude, with the highest alpha diversity in the milder areas of the McMurdo Dry Valleys (78{degrees}S) and the most southern areas (Durham Point, 85.5{degrees}S; Garden Spur, 84.5{degrees}S), and lowest in the especially arid and cold Darwin Area (~79.8{degrees}S). Furthermore, the specificity of mycobiont species towards their photobionts decreased under more severe climate conditions. The generalist lichen species Lecanora fuscobrunnea and Lecidea cancriformis were present in almost all habitats, but were dominant in climatically extreme areas. Carbonea vorticosa, Lecidella greenii and Rhizoplaca macleanii were confined to milder areas. In summary, the macroclimate is considered to be the main driver of species distribution, making certain species useful as bioindicators of climate conditions and, consequently, for detecting climate change.

ecology↗

bioRxiv2021

Floral scents of a deceptive plant are hyperdiverse and under population-specific phenotypic selection

Floral scent is a key mediator in plant-pollinator interactions; however, little is known to what extent intraspecific scent variation is shaped by phenotypic selection, with no information yet in deceptive plants. We recorded 291 scent compounds in deceptive moth fly-pollinated Arum maculatum from various populations north vs. south of the Alps, the highest number so far reported in a single plant species. Scent and fruit set differed between regions, and some, but not all differences in scent could be explained by differential phenotypic selection in northern vs. southern populations. Our study is the first to provide evidence that phenotypic selection is involved in shaping geographic patterns of floral scent in deceptive plants. The hyperdiverse scent of A. maculatum might result from the plants imitation of various brood substrates of its pollinators.

ecology↗

bioRxiv2019

Arabidopsis phytochelatin synthase 1, but not phytochelatin synthesis, functions in extracellular defense against multiple fungal pathogens

Phytochelatin synthase (PCS) is a key component of heavy metal detoxification in plants. PCS catalyzes both the synthesis of the peptide phytochelatin from glutathione as well as the degradation of glutathione conjugates via peptidase activity. Here, we describe a hitherto uncharacterized role for PCS in disease resistance against plant pathogenic fungi. The pen4 mutant, which is allelic to cadmium insensitive 1 (cad1/pcs1) mutants, was recovered from a screen for Arabidopsis mutants with reduced resistance to the non-adapted barley fungal pathogen, Blumeria graminis f. sp. hordei. PCS1, which is found in the cytoplasm of cells of healthy plants, translocates upon pathogen attack and colocalizes with the PEN2 myrosinase on the surface of immobilized mitochondria. pcs1 and pen2 mutant plants exhibit a similar metabolic defect in the accumulation of pathogen-inducible indole glucosinolate-derived compounds, suggesting that PEN2 and PCS1 act in the same metabolic pathway. The function of PCS1 in this pathway is independent of phytochelatin synthesis and deglycination of glutathione conjugates, as catalytic-site mutants of PCS1 are still functional in indole glucosinolate metabolism. In uncovering a previously unknown function for PCS1, we reveal this enzyme to be a moonlighting protein important for plant responses to both biotic and abiotic stresses.

plant biology↗