Biology subjects

Rodrigues, V.

Publications and source records attributed to Rodrigues, V..

8 recordsLinked to original sources

bioRxiv2026

Galleria mellonella as a novel invertebrate model for studying Ehrlichia ruminantium pathogenesis and host-pathogen interactions

Ehrlichia ruminantium, the causative agent of heartwater disease, is an obligate intracellular bacterium that poses significant economic threats to livestock production in endemic regions. Current research models present substantial ethical, logistical, and economic constraints, particularly for studying host-pathogen interactions within arthropod vectors. Here we establish Galleria mellonella larvae as a tractable invertebrate infection model for Ehrlichia ruminantium, enabling experimental investigation of pathogen persistence and host-pathogen interactions in an arthropod system. Following infection, G. mellonella proved susceptible to E. ruminantium with moderate mortality and remarkable bacterial persistence. Using rhodamine-labeled bacteria and fluorescence microscopy, we tracked bacterial dissemination from injection sites to systemic distribution in characteristic segmental patterns throughout the larval body. Critically, we confirmed intracellular localization of E. ruminantium within hemocytes, the primary immune cells of G. mellonella. Quantitative PCR analysis revealed stable bacterial loads over the study period, indicating bacterial persistence within the host. These findings demonstrate that E. ruminantium can hijack the innate immune system of G. mellonella, similar to its behavior in natural hosts. The segmental bacterial distribution suggests exploitation of hemolymph circulation and sessile hemocyte populations, providing new insights into potential mechanisms of pathogen persistence. This model offers significant advantages: ethical acceptability, cost-effectiveness, experimental tractability, and compatibility with high-throughput screening approaches. The G. mellonella system represents a valuable complement to existing mammalian models and provides a unique platform for investigating arthropod-specific aspects of E. ruminantium biology, screening antimicrobial compounds, and understanding mechanisms of immune evasion that may inform strategies for heartwater disease control.

microbiology↗

bioRxiv2025

GAS7 coordinates the activity of RAC1 and CDC42 to enhance macropinocytic surveillance and suppress motility in macrophages

Fluid uptake by macropinocytosis is central for the sentinel function of macrophages. However, how the actin network is organized at the cell surface to drive persistent membrane ruffling and macropinocytic cup formation remains unclear. Due to the high curvature of dorsal ruffles, we asked whether membrane curvature-associated proteins, such as those containing F-BAR domains, participate in this process. Here, leveraging public immune transcriptomic profiling and functional assays, we identify Growth Arrest Specific-7 (GAS7) as an F-BAR protein required for membrane ruffling and macropinocytosis in primary human monocyte-derived macrophages. GAS7 is enriched at dorsal ruffles and associates with actin regulators, sustaining high levels of active RAC1, to promote ruffle assembly. Moreover, GAS7 restrains the activation of CDC42, a GTPase linked to filopodia formation, cell motility and invasion. As a result, loss of GAS7 reduces ruffling and fluid uptake and instead promotes a phenotype characterized by increased motility, prominent filopodia, altered podosome dynamics, as well as an extracellular matrix-remodelling transcriptional program. Finally, we demonstrate that GAS7 enhances innate surveillance by augmenting the inflammatory response to muramyl dipeptide (NOD2 ligand) and by modulating the kinetics of TLR4 signalling to lipopolysaccharide. Thus, by coordinating RAC1 and CDC42 activities, GAS7 promotes macropinocytic uptake and enhances the macrophage responsiveness to microbial cues, while suppressing a more motile and invasive program.

cell biology↗

bioRxiv2025

A transient mechanical crosstalk between Adherens Junctions and Focal Adhesions sustains cell proliferation in premalignant breast epithelial cells

The weakening of Adherens Junctions (AJs) and Focal Adhesions (FAs) adhesiveness has been proposed to enable the initiation and progression of several types of cancer. Here we report that, prior to disassembling AJs and acquiring malignant traits, premalignant mammary epithelial cells overactivating the Src proto-oncogene transiently enhance tensile forces at both AJs and FAs, thereby gaining a proliferative advantage. We show that AJs and FAs are transiently under higher tensile forces in premalignant Src-activated cells. Cells unable to increase tensile forces at FAs by knocking down PXN or inhibiting FAK activity fail to transiently build up tensile force at AJs and to grow. Conversely, preventing AJ strengthening using EGTA or small interference RNA against P-cadherin suppresses the transient increase in tensile forces at FAs, EGFR-ERK and MRTF-A-SRF activation and cell proliferation. Moreover, knocking down E-cadherin, the sole classical cadherin in Drosophila, inhibits Src-induce tissue overgrowth in vivo. Thus, prior to the loss of cell-cell and cell-matrix adhesiveness, strengthening of AJs and FAs may be an essential early step for mammary cells to gain a proliferative advantage and establish the mechanical and signaling conditions necessary for subsequent malignant transformation. Statement of SignificanceAlthough the weakening of cell-cell and cell-matrix adhesiveness is commonly associated with cancer initiation and progression, our findings reveal that an initial increase in adhesiveness enables proliferation and malignant progression.

cancer biology↗

bioRxiv2025

Prevalence of Heartwater in Guadeloupe (2024): Stable endemicity and Evidence of Spread to Les Saintes

Heartwater is a tick-borne disease caused by Ehrlichia ruminantium and remains endemic in the Guadeloupe archipelago. While previous data from 1989 suggested a seroprevalence of 22% in cattle, no updated figures were available for livestock, and certain islands such as Les Saintes were considered disease-free. This study aimed to update the sero-epidemiological status of heartwater in cattle and goats across Guadeloupe in 2024. Blood samples were collected from 261 cattle and 135 goats from all islands. Serological testing using the MAP1B ELISA was performed, and true seroprevalence was estimated using a simplified stochastic model that accounts for test sensitivity, specificity, and sample uncertainty. The mean cattle seroprevalence was estimated at 28% (95% CI [22-34%]), with no significant differences between Basse-Terre, Grande-Terre, and Marie-Galante. Importantly, seropositive goats were identified on all major islands, including Les Saintes. Our findings suggest that heartwater remains endemically stable in Guadeloupe since the 1980s and reveal for the first time its serological presence in Les Saintes. These results emphasize the importance of strengthening regional surveillance systems, including veterinary reporting, serological monitoring, and coordinated efforts to prevent further spread across the Caribbean.

immunology↗

bioRxiv2025

Dissection and internal observation of the giant tropical bont tick Amblyomma variegatum

Ticks are parasites arthropods that feed on the blood of animals and humans, serving as vectors responsible for numerous diseases worldwide. The tropical bont tick, Amblyomma variegatum, is the primary vector of Ehrlichia ruminantium, the causative agent of heartwater, endemic in sub-Saharan and southern Africa and several Caribbean islands. Due to their economic and public health impacts, A. variegatum and heartwater have been classified as high-consequence foreign animal diseases and pests by the United States Department of Agricultures Animal and Plant Health Inspection Service. Understanding the ecology and biology of A. variegatum is thus crucial to describe the tick-bacteria interactions and thus developing effective control methods. A key initial step in this endeavor involves the precise isolation and detailed functional characterization of tick organs to better understand vector competence. However, complete anatomical descriptions of internal structures within the Amblyomma genus remain scarce. In this study, we present a thorough characterization of the internal anatomy of A. variegatum. By accurately identifying and mapping organs of interest, we highlight notable morphological differences from other tick species. This work not only enhances our understanding of A. variegatum anatomy but also serves as a crucial resource for investigating tick-borne disease transmission. Ultimately, these findings support the development of targeted vector control strategies, applicable to both endemic and non-endemic regions.

zoology↗

bioRxiv2024

A human homolog of SIR2 antiphage proteins mediates immunity via the TLR pathway

The full extent of immune system conservation between prokaryotes and eukaryotes is unknown. However, recent research supports that a subset of bacterial antiphage proteins is conserved in eukaryotes and likely gave rise to key actors of mammalian immunity. Here, we show that the SIR2 protein domain, present in bacterial antiphage systems, plays a role in eukaryotic innate immunity. Through phylogenetic analysis, we identify SIRanc, a human protein with a SIRim domain (subtype of SIR2). We demonstrate that SIRanc plays a pivotal role in the animal toll-like receptor (TLR) pathway of innate immunity by mediating the transcriptional upregulation of proinflammatory genes downstream of TLR stimulation. This depends on the enzymatic activity of SIRanc, which degrades NAD+, a central cellular metabolite. Finally, we show that proteins with a SIRim domain are diverse and widespread, detected in 19% of eukaryotic genomes, with SIRanc representing one of the five sirim lineages. This work opens avenues of research on the potential role of eukaryotic SIRim proteins in immunity, as well as on the involvement of SIRanc in human pathology.

immunology↗

bioRxiv2022

The m6A writer FIONA1 methylates the 3'UTR of FLC and controls flowering in Arabidopsis

Adenosine bases of RNA can be transiently modified by the deposition of a methyl-group to form N6-methyladenosine (m6A). This adenosine-methylation is an ancient process and the enzymes involved are evolutionary highly conserved. A genetic screen designed to identified suppressors of late flowering transgenic Arabidopsis plants overexpressing the miP1a microProtein yielded a new allele of the FIONA1 (FIO1) m6A-methyltransferase. To characterize the early flowering phenotype of fio1 mutant plants we employed an integrative approach of mRNA-seq, Nanopore direct RNA-sequencing and meRIP-seq to identify differentially expressed transcripts as well as differentially methylated mRNAs. We provide evidence that FIO1 is the elusive methylase responsible for the 3-end methylation of the FLOWERING LOCUS C (FLC) transcript. Furthermore, our genetic and biochemical data suggest that 3-methylation stabilizes FLC mRNAs and non-methylated FLC is a target for rapid degradation.

plant biology↗

bioRxiv2022

Release of HIV-1 particles from the viral compartment in macrophages requires an associated cytoskeleton and is driven by mechanical constraints

A defining feature of HIV-1 replication in macrophages is that viral assembly occurs at the limiting membrane of a compartment often named VCC (virus-containing compartments) that is connected to the extracellular medium. The newly formed viral progeny pinches of the membrane and accumulates in the lumen of the VCC. While HIV budding has been extensively studied, very little is known about how viral particles present in the lumen of VCC are released in the extracellular medium. Here we show that the actin dynamics are critical for this process by combining ultrastructural analyses, time-lapse microscopy and perturbations of the actin cytoskeleton. We found that jasplakinolide, which stabilizes actin fibres, inhibited viral release from HIV-1-infected macrophages. Furthermore, in jasplakinolide-treated macrophages, VCC became scattered and no longer co-localized with the integrin CD18, nor the phosphorylated form of the focal adhesion kinase PYK2. Inhibition of PYK2 activity in infected macrophages promoted intracellular retention of viral particles in VCC that were no longer connected to the plasma membrane. Finally, we stimulated the rapid release of viral particles from the VCC by subjecting infected macrophages to frustrated phagocytosis. As macrophages spread on IgG-coated glass surfaces, VCC rapidly migrated to the basal membrane and released their viral content in the extracellular medium, which required their association with CD18 and the actin cytoskeleton. These results highlight that VCC trafficking and virus release are intimately linked to the reorganization of the macrophage actin cytoskeleton in response to external physical cues, suggesting that it might be regulated in tissues by the mechanical stress to which these cells are exposed.

cell biology↗