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Biology subjects

Marino, G.

Publications and source records attributed to Marino, G..

14 recordsLinked to original sources

Distinct roles for DNA- and RNA-sensing immune pathways in control of genital HSV-2 infection and restriction of neuroinvasion

Early control of viral infection is thought to rely on pattern-recognition receptors (PRRs) that induce interferons (IFNs) and leukocyte recruitment, but how distinct PRRs coordinate mucosal antiviral defense remains unclear. We show that both the DNA-sensing cGAS-STING pathway and the RNA-sensing RLR-MAVS pathway are required for protection upon genital herpes simplex virus type 2 (HSV-2) infection. cGAS deficiency increased infection-induced pathology in both epithelial and submucosal compartments, whereas MAVS deficiency primarily affected the epithelium. Spatial proteomics and regional transcriptomics revealed that cGAS was essential for early epithelial TBK1 activation, expression of IFN-stimulated genes and recruitment and activation of myeloid and lymphoid cells to the epithelium. While MAVS was essential for full TBK1 activation it had limited impact on the induced IFN response. However, MAVS sustained basal epithelial expression of the antiviral factors IFITM1 and 3, which exert antiviral activity against HSV-2. Notably, cGAS deficiency impaired submucosal IFN responses and enabled viral spread into this tissue, enabling infection of intervening neurons and dissemination to the central nervous system. These findings define coordinated, compartment-specific innate defense against infections.

immunology↗

Au_Sus: A tiered consensus census of human autophagy genes

Autophagy is a critical cellular process, yet its genomic definition remains inconsistent across digital repositories. This lack of standardisation hinders reproducibility in high-throughput studies and clinical research. Here, we present Au_Sus, a high-confidence human autophagy census established through a frequency-based majority consensus of seven primary databases and literature sources. After rigorous manual curation and nomenclature standardisation, we defined a tiered framework: Maxim_Au (2,581 genes), Au_Sus (the 201-gene core consensus), and Minim_Au (77 universal genes). Functional enrichment and protein-protein interaction analysis confirm that Au_Sus captures a highly integrated and purified autophagic machinery, with significant associations to neurodegeneration and oncology. Furthermore, an analysis of nearly 100 published cancer gene signatures revealed profound functional dilution, with 60% of signature genes absent from our consensus. These findings suggest that many of these models incorporate peripheral stress markers rather than core autophagic effectors. Hence, Au_Sus (freely accessible at ausis.uniovi.es) provides a reliable, ready-to-use benchmark to standardise the study of autophagy in health and disease.

cell biology↗

Bayesian neural networks enable inference of complex phylodynamic processes

Phylogenetic branching patterns carry essential information about population and diversification dynamic processes, including speciation, extinction, and epidemiological transmission. Phylodynamic models offer a rigorous mathematical framework for quantifying these dynamics from phylogenetic trees. Extensions of these models enable the incorporation of external covariates as predictors of phylodynamic parameters, for instance allowing us to link traits or environmental variables with changes in speciation, extinction, or transmission rates. However, the dependencies between predictors and phylodynamic parameters are typically restricted to linear, additive effects and may thus fail to capture complex, potentially non-linear relationships underlying evolutionary dynamics. To address this limitation, we propose a new framework, BELLA, which leverages unsupervised Bayesian neural networks (BNNs) to flexibly model functional relationships between key phylodynamic parameters and a broad set of predictors, including categorical traits, quantitative variables, and time series data. Based on these covariates, the BNN weights are estimated through Markov chain Monte Carlo and can be inferred jointly with the phylogenetic tree topology and branching times, obtained directly from sequence alignment data. Using extensive simulations, we demonstrate that this approach accurately recovers predictor-parameter relationships, mitigates overfitting, and remains robust across both macroevolutionary and epidemiological contexts. By incorporating tools from explainable artificial intelligence, we further show that our framework reliably identifies the most influential predictors and yields interpretable descriptions of their impact on phylodynamic rates. Finally, we apply our method to two empirical analyses: linking SARS-CoV-2 migration dynamics with travel data during its early spread in Europe, and inferring trait and time-dependent speciation and extinction rates in the Cenozoic diversification of platyrrhines. Our unsupervised BNN framework substantially expands the capabilities of phylodynamic inference providing a powerful and flexible approach to model complex macroevolutionary and epidemiological processes. Significance statementUnderstanding how species diversify or pathogens spread requires linking phylogenetic trees to biological traits and environmental characteristics. We present BELLA, an unsupervised Bayesian neural network that learns complex, nonlinear links between predictors--such as traits or environmental variables--and key rates such as speciation, extinction, transmission, and migration directly from sequence data--which in turn inform on the phylogenetic relationships. BELLA does not require any training data, enabling inferences that were previously not tractable. Across simulated epidemics and macroevolutionary histories, BELLA is more accurate than state-of-the-art linear models relating predictors and rates, while remaining interpretable through explainable artificial intelligence. Empirical applications in epidemiology reveal nonlinear effects of travel among countries on the early spread of SARS-CoV-2 in Europe, while a macroevolutionary application shows selective extinction events in the diversifcation history of New World monkeys. The BELLA framework expands the reach of phylodynamics in biology and public health.

evolutionary biology↗

Key molecular and cellular events of table olive fruit abscission zone formation during natural maturation and after ethephon treatment

Abscission zones are specialized cell layers that enable organ detachment, yet their morphology and regulation vary among species. Olive (Olea europaea), a non-climacteric fruit, requires high concentrations of ethylene-releasing compounds to reduce fruit removal force (FRF) for mechanical harvest. To elucidate mechanisms of fruit abscission zone (FAZ) development in olive, we integrated physiological, transcriptomic, and cellular analyses during natural maturation and after ethephon treatment. Olive fruits emitted low ethylene at color transition, coinciding with declining FRF, and application of 1500 ppm ethephon reduced FRF after one week. Transcriptome analyses of FAZ tissues, using a mesocarp-subtraction strategy to isolate FAZ-specific responses, identified 733 FAZ-specific genes shared between natural maturation and ethephon treatments, including genes of {beta}-1,3-glucanases, pectate lyases, and the phenylpropanoid pathway. Microscopy revealed lignification and alkalization of FAZ, accompanied by reduced low-methylesterified homogalacturonan and non-fucosylated xyloglucan. Increased glucanase activity and reduction in plasmodesmata callose likely facilitate cellular communication during abscission. Further, this study provides the first evidence of FAZ alkalization and implicates transporter upregulation in pH regulation preceding abscission. Our findings advance understanding of abscission biology in non-climacteric fruits and delineate conserved features of FAZ development. Summary StatementIntegrated physiological, transcriptomic, and cellular analyses reveal that fruit abscission zone development in olive involves lignification, alkalization, and coordinated cell wall remodeling, identifying {beta}-1,3-glucanases, pH-regulating transporters, and pectate lyases that facilitate cell separation in a non-climacteric fruit.

plant biology↗

ATG4D loss leads to late-onset cardiomyopathy and stress-induced heart failure in mice, and its repression marks maladaptive cardiac remodeling in humans

In the last years, autophagy has emerged as an essential pathway for most cellular functions. Basal autophagy plays a protective role as a quality control mechanism by which damaged or noxious cellular components are degraded and cellular organelles are periodically renewed. Moreover, autophagic activity can be increased in situations of cellular stress, including nutrient or growth factor deprivation, hypoxia, reactive oxygen species, DNA damage, or the presence of intracellular pathogens. Normally, induction of autophagy is protective, although in some circumstances, such as conditions of hemodynamic stress, autophagosome accumulation upon autophagy induction can be a maladaptive process. The deficiency of the autophagic protease ATG4D in mice leads to the accumulation of cellular autophagosomes in most tissues, including the heart. Here, we show that the increased autophagosome content of atg4d-/- mice is linked to the development of late-onset cardiomyopathy and to increased susceptibility to heart failure induced by transverse aortic constriction. Furthermore, we report the existence of human ATG4D variants associated with cardiovascular pathologies and also that ATG4D expression is reduced in human obstructive hypertrophic cardiomyopathy and dilated cardiomyopathy, which highlights a conserved cardio-protective role of the ATG4D protease.

pathology↗

The functional divergence of plant ESCRT components TOL3, SNF7.1, and VPS4 during salt stress response

Salinity, a major abiotic stressor, impacts protein trafficking and alters transport routes. The endosomal sorting complex required for transport (ESCRT)is a key player for membrane protein sorting and is increasingly recognized as pivotal element in abiotic stress responses. However, the functional role of ESCRT proteins in cereal crops under salt stress remains unexplored. Using the as near as possible to nature DRD-BIBLOX (Dark-Root device Brick Black Box), barley (Hordeum vulgare, L.), although relatively salt-tolerant, exhibits impaired germination, seedling growth, and an altered root system architecture (RSA) under high salinity. Shotgun proteomic analysis revealed spatio-temporal regulation of ESCRT proteins in shoots and roots during seedling development. We identified HvSNF7.1 and HvVPS4 as critical, but differently acting modulators of salt stress responses in barley tissues. In this study we further reveal novel functions of ESCRT-0-like TOM-1-like proteins (TOLs): Previously described as gatekeepers of degradative protein sorting and as regulators of abscisic acid signaling, growth, and heat stress responses in Arabidopsis (Arabidopsis thaliana), HvTOLs had not been characterized for their role in salinity adaptation during plant development. Functional analyses demonstrated lethality in Hvtoldouble and Hvtolquadruple mutants under standard growth conditions. In Arabidopsis, loss of AtTOL3 alone impaired germination and root growth under salt stress, underscoring the essential role of individual TOL proteins in the spatio-temporal regulation of abiotic stress adaptation. These findings highlight ESCRT-mediated trafficking as a key determinant of cereal resilience under saline conditions and provide a molecular framework for improving crop stress tolerance.

plant biology↗

Mrx6 binds the Lon protease Pim1 N-terminal domain to confer selective substrate specificity and regulate mtDNA copy number

Mitochondrial DNA (mtDNA) copy number regulation remains incompletely understood, despite its importance in cellular function. In Saccharomyces cerevisiae, Mrx6 belongs to the Pet20-domain-containing protein family, consisting of Mrx6, Pet20, and Sue1. Notably, absence of Mrx6 leads to increased mtDNA copy number. Here, we identify the C-terminus of Mrx6 as essential for its stability and interaction with the mitochondrial matrix protein Mam33. Deletion of Mam33 mimics the effect of Mrx6 loss, resulting in elevated mtDNA copy number. Bioinformatics, mutational analyses, and immunoprecipitation studies reveal that a subcomplex of Mam33 and Mrx6 trimers interacts with the substrate recognition domain of the conserved mitochondrial Lon protease Pim1 through a bipartite motif in the Pet20 domain of Mrx6. Loss of Mrx6, its paralog Pet20, Mam33, or mutations disrupting the interaction between Mrx6 and Pim1 stabilize key proteins required for mtDNA maintenance, the RNA polymerase Rpo41 and the HMG-box-containing protein Cim1. We propose that Mrx6, alongside Pet20 and Mam33, regulates mtDNA copy number by modulating substrate degradation through Pim1. Additionally, Mrx6 loss alters Cim1s function, preventing the detrimental effect on mtDNA maintenance observed upon Cim1 overexpression. The presence of three Pet20-domain proteins in yeast implies broader roles of Lon protease substrate recognition beyond mtDNA regulation.

cell biology↗

Reduced systemic autophagy by simultaneous loss of ATG4B, ATG4C and ATG4D leads to accelerated aging in mice

Autophagy is an essential catabolic pathway that safeguards cellular and tissue homeostasis, yet the systemic consequences of its impairment in mammals remain poorly defined because complete autophagy ablation is embryonic or perinatal lethal. Here, we generate ATG4A-only mice, a model in which ATG4A is the sole remaining ATG4 protease due to combined ATG4B/C/D deletion. Through comprehensive biochemical and cellular analyses, we delineate the in vivo substrate specificity of ATG4A and demonstrate that it sustains only minimal ATG8 priming, uncovering a previously unrecognized functional asymmetry within the mammalian ATG4-ATG8 system. ATG4A-only mice exhibit a profound but incomplete whole-body autophagy deficiency that disrupts multiple organ systems and triggers a premature aging syndrome marked by increased DNA damage, systemic senescence, metabolic dysfunction, and dramatically shortened lifespan. Integrating these findings with comparisons to additional ATG4-deficient models, we show that organismal longevity scales with residual autophagic competence. Together, our work reveals how graded reductions in autophagy integrity influence tissue fitness and aging, establishing autophagic capacity as a key determinant of mammalian lifespan.

cell biology↗

In a nutshell: pistachio genome and kernel development

Pistachio is a sustainable nut crop with exceptional climate resilience and nutritional value. To advance pistachios as a future food source and a model system for hard-shelled fruits, we generated a chromosome-scale reference genome of the most widely grown pistachio cultivar (Pistacia vera Kerman) and a spatiotemporal developmental study of the hull, shell, and kernel. Our study defined four distinct stages of pistachio growth and maturation by integrating tissue-level physiological and molecular data from thousands of nuts across twenty-four time points over three growing seasons. Transcriptional and metabolic changes in the kernel elucidate molecular pathways governing nutritional quality, such as the accumulation of unsaturated fatty acids, which are vital for shelf-life and dietary value. This work yields new knowledge and resources that will inform other woody crops and facilitate further improvement of pistachio as a globally significant, sustainable, and nutritious crop.

plant biology↗

A pgr5 suppressor screen uncovers a distinct mechanism safeguarding the cytochrome b6f complex from damage through PGR5

PROTON GRADIENT REGULATION5 (PGR5) is thought to promote cyclic electron flow (CEF) and its deficiency causes increased photosensitivity of photosystem I (PSI), leading to lethality under fluctuating light (FL). By screening for suppressor mutations that rescue FL lethality of pgr5 plants, we identified a portfolio of mutations affecting 12 photosynthesis-related proteins. Six are required for proper PSII function, one (CcdA) promotes cytochrome (cyt) b6f assembly, and another (PAA1) provides plastocyanin with its copper cofactor. Two other mutations are associated with the chloroplast FBPase cFBP1. This, together with targeted knockout of other genes in the pgr5 background, suggests three pathways to restore FL viability: (i) reduced electron flow to PSI due to defects in PSII, cyt b6f or plastocyanin but not PSI, (ii) increased electron flow from PSI due to inactivation of ACHT2, a regulator of cFBP1 activity, and (iii) hyperactivity of the NDH-dependent CEF due to inactivation of cFBP1. The remaining two suppressor mutations affected the cyt b6f complex. PFSC1 controls cyt b6f accumulation at early developmental stages. DEIP1/NTA1, previously suggested to be essential for cyt b6f assembly, appears to protect cyt b6f from deleterious effects of PGR5, since plants lacking both DEIP1/NTA1 and PGR5 are viable and accumulate cyt b6f.

plant biology↗

A bench-top dark-root device built with LEGO bricks enables a non-invasive plant root development analysis in soil conditions mirroring nature

Roots are the hidden parts of plants, anchoring their above ground counterparts in the soil. They are responsible for water and nutrient uptake, as well as for interacting with biotic and abiotic factors in the soil. The root system architecture (RSA) and its plasticity are crucial for resource acquisition and consequently correlate with plant performance, while being highly dependent on the surrounding environment, such as soil properties and therefore environmental conditions. Thus, especially for crop plants and regarding agricultural challenges, it is essential to perform molecular and phenotypic analyses of the root system under conditions as near as possible to nature (#asnearaspossibletonature). To prevent root illumination during experimental procedures, which would heavily affect root development, dark-root (D-Root) devices (DRDs) have been developed. In this article, we describe the construction and different applications of a sustainable, affordable, flexible, and easy to assemble open-hardware bench-top LEGO(R) DRD, the DRD-BIBLOX (Brick Black Box). The DRD-BIBLOX consists of one or more 3D-printed rhizoboxes which can be filled with soil, while still providing root visibility. The rhizoboxes sit in a scaffold of secondhand LEGO(R) bricks, which allows root development in the dark as well as non-invasive root-tracking with an infrared (IR) camera and an IR light emitting diode (LED) cluster. Proteomic analyses confirmed significant effects of root illumination on barley root and shoot proteome. Additionally, we confirmed the significant effect of root illumination on barley root and shoot phenotypes. Our data therefore reinforces the importance of the application of field conditions in the lab and the value of our novel device, the DRD-BIBLOX. We further provide a DRD-BIBLOX application spectrum, spanning from investigating a variety of plant species and soil conditions as well as simulating different environmental conditions and stresses, to proteomic and phenotypic analyses, including early root tracking in the dark.

plant biology↗

Ambra1 haploinsufficiency results in metabolic alterations and exacerbates age-associated retinal degeneration

Autophagy is a key process in the maintenance of cellular homeostasis. The age-dependent decline in retinal autophagy has been associated with photoreceptor degeneration. Retinal dysfunction can also result from damage to the retinal pigment epithelium (RPE), as the RPE-retina constitutes an important metabolic ecosystem that must be finely tuned to preserve visual function. While studies of mice lacking essential autophagy genes have revealed a predisposition to retinal degeneration, the consequences of a moderate reduction in autophagy, similar to that which occurs during physiological aging, remain unclear. Here, we describe a retinal phenotype consistent with accelerated aging in mice carrying a haploinsufficiency for Ambra1, a pro-autophagic gene. These mice show protein aggregation in the retina and RPE, metabolic underperformance, and premature vision loss. Moreover, Ambra1+/gt mice are more prone to retinal degeneration after RPE stress. These findings indicate that autophagy provides crucial support to RPE-retinal metabolism and protects the retina against stress and physiological aging.

cell biology↗

Microglial phagocytosis dysfunction during stroke is prevented by rapamycin

Microglial phagocytosis is rapidly emerging as a therapeutic target in neurodegenerative and neurological disorders. An efficient removal of cellular debris is necessary to prevent buildup damage of neighbor neurons and the development of an inflammatory response. As the brain professional phagocytes, microglia are equipped with an array of mechanisms that enable them to recognize and degrade several types of cargo, including neurons undergoing apoptotic cell death. While microglia are very competent phagocytes of apoptotic cells under physiological conditions, here we report their dysfunction in mouse and monkey (Macaca fascicularis and Callithrix jacchus) models of stroke by transient occlusion of the medial cerebral artery (tMCAo). The impairment of both engulfment and degradation was related to energy depletion triggered by oxygen and nutrients deprivation (OND), which led to reduced process motility, lysosomal depletion, and the induction of a protective autophagy response in microglia. Basal autophagy, which is in charge of removing and recycling intracellular elements, was critical to maintain microglial physiology, including survival and phagocytosis, as we determined both in vivo and in vitro using knock-out models of autophagy genes and the autophagy inhibitor MRT68921. Notably, the autophagy inducer rapamycin partially prevented the phagocytosis impairment induced by tMCAo in vivo but not by OND in vitro. These results suggest a more complex role of microglia in stroke than previously acknowledged, classically related to the inflammatory response. In contrast, here we demonstrate the impairment of apoptotic cell phagocytosis, a microglial function critical for brain recovery. We propose that phagocytosis is a therapeutic target yet to be explored and provide evidence that it can be modulated in vivo using rapamycin, setting the stage for future therapies for stroke patients.

neuroscience↗

AtCGL160 recruits chloroplast coupling factor 1

ATP synthases couple the generation of chemical energy to a transmembrane electro-chemical potential. Like ATP synthases in bacteria and mitochondria, chloroplast ATP synthases consist of a membrane-spanning (CFO) and a soluble coupling factor (CF1). Accessory factors facilitate subunit production and orchestrate the assembly of the functional CF1-CFO complex. It was previously shown that the accessory factor CGL160 promotes the formation of plant CFO and performs a similar function in the assembly of its c-ring to that of the distantly related bacterial Atp1/UncI protein. In this study, we show that the N-terminal portion of CGL160 (AtCGL160N), which is specific to the green lineage, is required for late steps in CF1-CFO assembly in Arabidopsis thaliana. In plants that lacked this stroma-exposed domain, photosynthesis was impaired, and amounts of CF1-CFO were reduced to about 65% of the wild-type level. Loss of AtCGL160N did not perturb c-ring formation, but led to a 10-fold increase in the numbers of CF1 sub-complexes in the stroma relative to the wild type and the CF1 assembly mutant atcgld11-1. Co-immunoprecipitation and protein crosslinking assays revealed an association of AtCGL160 with CF1 subunits. Yeast two-hybrid assays localized the interaction to a stretch of AtCGL160N that binds to the thylakoid-proximal domain of CF1-{beta} that includes the conserved DELSEED motif. We therefore propose that AtCGL160 has acquired an additional function in the recruitment of soluble CF1 to a membrane-integral CFO sub-complex, which is critical for the modulation of CF1-CFO activity and photosynthesis in chloroplasts.

plant biology↗