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Biology subjects

Lopes, J.

Publications and source records attributed to Lopes, J..

7 recordsLinked to original sources

A major life-history locus underlies genotype-by-environment variation in growth across water temperatures

Rapid environmental change is exposing organisms to conditions that do not match those under which they evolved, making it increasingly important to understand how genetic variation shapes phenotypic responses to environmental stress. Since most phenotypic traits arise from interactions between genetic variation and the environments experienced throughout an organisms lifetime, understanding the genetic architecture of these interactions is central to predicting how populations will respond to novel environments. While genotype-by-environment interactions (GxE) are well studied in quantitative genetics, identifying specific loci that contribute to environmentally dependent trait expression remains rare. Salmonids already exhibit a wide portfolio of plastic life-history strategies, reflecting adaptation to highly heterogeneous environments, yet it remains unclear whether known major-effect loci involved in life-history regulation also contribute to variation in plastic responses to environmental change. One such major-effect locus is the transcription factor six6, which has been repeatedly associated with variation in age at maturity across multiple populations of rainbow trout (Oncorhynchus mykiss). Since maturation timing is closely linked to growth trajectories and patterns of energetic allocation during early development, allelic variation at this locus may also influence growth responses to warming conditions. Here, we test this hypothesis using a common-garden experiment in which 6 months old juvenile rainbow trout were reared under current and warming (+2{degrees}C) temperature regimes. By quantifying genotype-specific reaction norms across environments, we show that six6 genotype contributes to environmentally dependent variation in growth and body composition, with individuals heterozygous for the six6 locus showing a distinct and steeper response to warming relative to homozygotes. These findings provide evidence that a major life-history gene shapes plastic responses to thermal stress in juvenile rainbow trout, with novel implications for how standing genetic variation at in large-effect loci may influence population-level responses to climate warming.

evolutionary biology↗

PRO-FitS: a novel phenotypic assay to identify enhancers of proteostasis in C. elegans

The prevalence of neurodegenerative diseases (NDs) continues to rise with the aging of populations worldwide, representing a pressing need for the establishment of therapeutic strategies. Maintaining proteostasis is crucial for healthy aging, as the accumulation of misfolded and aggregated proteins is a key contributor to age-related cellular dysfunction and disease. This study introduces a novel phenotypic assay using Caenorhabditis elegans to screen for small molecule enhancers of proteostasis, aiming at mitigating the proteotoxic stress associated with NDs. This new methodology- PRO-FitS- uses C. elegans motor activity as a proxy for the PROteome Fitness State upon a noxious protein-denaturating stimulus, while allowing a fast and experimenter-free readout. We demonstrate the efficacy of the assay by validating the role of pharmacological mTOR inhibition and serotonergic signaling activation in reducing heat shock-induced proteotoxic damage at the whole-organism level. PRO-FitS will allow the identification of novel compounds that alleviate protein aggregation disorders, potentially revealing new pathways and cellular targets not previously implicated in proteotoxicity. Significance StatementNeurodegenerative diseases remain without effective cures, in part due to the lack of scalable methods to identify compounds that improve proteostasis. We developed PRO-FitS, a whole-organism, automated phenotypic assay in C. elegans that uses motor activity recovery as a proxy for proteome fitness after proteotoxic stress. This platform enables rapid, unbiased screening of small molecules and genetic modifiers, bridging the gap between cellular assays and complex animal models. By demonstrating the assays robustness in both wild-type and disease-relevant contexts, we establish PRO-FitS as a versatile tool for discovering therapeutic candidates and uncovering novel pathways relevant to protein aggregation disorders.

neuroscience↗

Dynamic ensembles of SARS-CoV-2 N-protein reveal head-to-head coiled-coil-driven oligomerization and phase separation

The SARS-CoV-2 nucleocapsid (N) protein is essential for the viral lifecycle, facilitating RNA packaging, replication, and host-cell interactions. Its ability to self-assemble and undergo liquid-liquid phase separation (LLPS) is critical for these functions but remains poorly understood. Using an integrated approach combining small-angle X-ray scattering (SAXS), nuclear magnetic resonance (NMR) spectroscopy, computational modeling, and biophysical assays, we uncover key mechanisms underpinning N-proteins dynamic self-assembly. We show that the N-proteins interdomain linker (IDL) contains a conserved coiled-coil (CC) motif that drives transient interactions between protein subunits, enabling the formation of progressively larger complexes at higher concentrations. SAXS analysis and ensemble modeling reveal that the IDL exists in a concentration-dependent equilibrium between monomeric, dimeric, and trimeric states. The CC motif facilitates parallel, head-to-head oligomerization of N-protein dimers, transitioning between compact (closed) and extended (open) configurations depending on the interaction network within the IDL. This linker-driven assembly modulates LLPS, impacting the size, stability, and dynamics of biomolecular condensates. Here, we present the most comprehensive conformational landscape analysis of the N-protein to date, providing a detailed model of its self-assembly and LLPS. Our findings highlight how the structural plasticity of the IDL and CC-mediated interactions are pivotal to its roles in the SARS-CoV-2 lifecycle.

biophysics↗

Socialization of Providencia stuartii enables resistance toenvironmental insults.

Providencia stuartii is a highly-social pathogen responsible for nosocomial chronic urinary tract infections. The bacterium indeed forms floating communities of cells (FCC) besides and prior-to canonical surface-attached biofilms (SAB). Within P. stuartii FCC, cells are riveted one to another owing to by self-interactions between its porins, viz. Omp-Pst1 and Omp-Pst2. In pathophysiological conditions, P. stuartii is principally exposed to high concentrations of urea, ammonia, bicarbonate, creatinine and to large variations of pH, questioning how these environmental cues affect socialization, and whether formation of SAB and FCC protects cells against those. Results from our investigations indicate that FCC and SAB can both form in the urinary tract, endowing cells with increased resistance and fitness. They additionally show that while Omp-Pst1 is the main gateway allowing penetration of urea, bicarbonate and ammonia into the periplasm, expression of Omp-Pst2 enables resistance to them.

microbiology↗

Epigenetic rewriting at centromeric DNA repeats leads to increased chromatin accessibility and chromosomal instability

Centromeric regions of human chromosomes contain large numbers of tandemly repeated -satellite sequences. These sequences are covered with constitutive heterochromatin which is enriched in trimethylation of histone H3 on lysine 9 (H3K9me3). Although well studied using artificial chromosomes and global perturbations, the contribution of this epigenetic mark to chromatin structure and genome stability remains poorly known in a more natural context. Using transcriptional activator-like effectors (TALEs) fused to a histone lysine demethylase (JMJD2B), we were able to reduce the level of H3K9me3 on the -satellites repeats of human chromosome 7. We show that the removal of H3K9me3 affects chromatin structure by increasing the accessibility of DNA repeats to the TALE protein. Tethering TALE-demethylase to centromeric repeats impairs the recruitment of HP1 and proteins of Chromosomal Passenger Complex (CPC) on this specific centromere. Finally, the epigenetic re-writing by the TALE-JMJD2B affects specifically the stability of chromosome 7 upon mitosis, highlighting the importance of H3K9me3 in centromere integrity and chromosome stability, mediated by the recruitment of HP1 and the CPC.

molecular biology↗

On the front line of Klebsiella pneumoniae surface structures understanding: establishment of Fourier Transform Infrared (FT-IR) spectroscopy as a capsule typing method

Genomics-based population analysis of multidrug resistant (MDR) Klebsiella pneumoniae (Kp) motivated a renewed interest on capsule (K) types given their importance as evolutionary and virulence markers of clinically relevant strains. However, there is a gap between genotypic based predictions and information on capsular polysaccharide structure and composition. We used molecular genotypic, comparative genomics, biochemical and phenotypic data on the cps locus to support the usefulness of Fourier-Transform Infrared (FT-IR) spectroscopy as a phenotypic approach for K-type characterization and identification. The approach was validated with a collection of representative MDR Kp isolates from main lineages/Clonal Groups (CGs) involved in local or nationwide epidemics in 6 European and South American countries. FT-IR-based K-type assignments were compared with those obtained by genotypic methods and WGS (cps operon), and further complemented with data on the polysaccharide composition and structure of known K-types. We demonstrate that our FT-IR-based spectroscopy approach can discriminate all 21 K-types identified with a resolution comparable (or even higher) to that provided by WGS, considered gold-standard methodology. Besides contributing to enlighten K-type diversity among a significant MDR Kp collection, the specific associations between certain K-types and Kp lineages identified in different geographic regions over time support the usefulness of our FT-IR-based approach for strain typing. Additionally, we demonstrate that FT-IR discriminatory ability is correlated with variation on the structure/composition of known K-types and, supported on WGS data, we were able to predict the sugar composition and chemical structure of new KL-types. Our data revealed an unprecedent resolution at a quick and low-cost rate of Kp K-types at the phenotypic level. Our FT-IR spectroscopy-based approach might be extremely useful not only as a cost-effective Kp typing tool, but also to improve our understanding on sugar-based coating structures of high relevance for strain evolution and host adaptation.

developmental biology↗

The GRASP domain in Golgi Reassembly and Stacking Proteins: differences and similarities between lower and higher Eukaryotes

The Golgi complex is part of the endomembrane system and is responsible for receiving transport cargos from the endoplasmic reticulum and for sorting and targeting them to their final destination. To perform its function in higher eukaryotic cells, the Golgi needs to be correctly assembled as a flatted membrane sandwich kept together by a protein matrix. The correct mechanism controlling the Golgi cisternae assembly is not yet known, but it is already accepted that the Golgi Reassembly and Stacking Protein (GRASP) is a main component of the Golgi protein matrix. Unlike mammalian cells, which have two GRASP genes, lower eukaryotes present only one gene and distinct Golgi cisternae assembly. In this study, we performed a set of biophysical studies to get insights on both human GRASP55 and GRASP65 and compare them with GRASPs from lower eukaryotes (S. cerevisiae and C. neoformans). Our data suggest that both human GRASPs are essentially different from each other and GRASP65 is more similar to the subgroup of GRASPs from lower eukaryotes. GRASP55 is present mainly in the Golgi medial and trans faces, which are absent in both funguses, while GRASP65 is located in the cis-Golgi. We suggest that the GRASP65 gene is more ancient and the paralogue GRASP55 might have appeared latter in evolution, together with the medial and trans Golgi faces in mammalians.

biophysics↗