Biology subjects

Ribeiro, D. M.

Publications and source records attributed to Ribeiro, D. M..

11 recordsLinked to original sources

bioRxiv2025

Cell-specific regulation of TCA cycle and cell division related genes at roots following aluminium exposure

Aluminium (Al) toxicity is a critical factor that limits plant growth in acidic soils (pH < 5.5) worldwide. Intracellular Al can bind to various biomolecules and significantly affect gene expression, protein biosynthesis, and cell membrane integrity, ultimately inhibiting nutrients and water uptake. This leads to the arrest of root cell divisions and growth. Organic acids (OAs) produced in mitochondrial-related reactions are strongly linked to Al tolerance, as they bind to Al and neutralize its toxic effects. Recently, the manipulation of root cell divisions through DNA checkpoints has been proposed as an alternative to increase Al tolerance. In this context, molecular interactions between these two mechanisms could potentially enhance plant tolerance to Al toxicity. Here, we investigate this topic using a translatome approach, which enables specific analyses of root cell types (namely, whole root, the quiescent centre region, and the cortical region). This approach offers a promising tool for unravelling how mitochondrial metabolism, particularly the TCA cycle, and the DNA damage response (DDR) pathway are interconnected in Al-mitigation process. Our results revealed that the TCA cycle is induced in differentiated cells, with no significant alteration in meristematic cells, while genes associated with cell cycle progression and DNA checkpoints show differential expression across the distinct cell types. Collectively, our data contribute to understand cell-specific internal detoxification mechanism related to Al tolerance in plants. This knowledge could assist crop breeding by providing a better understanding of the cellular physiological responses to Al toxicity.

plant biology↗

bioRxiv2022

Accurate rare variant phasing of whole-genome and whole-exome sequencing data in the UK Biobank

The UK Biobank performed whole-genome sequencing (WGS) and whole-exome sequencing (WES) across hundreds of thousands of individuals, allowing researchers to study the effects of both common and rare variants. Haplotype phasing distinguishes the two inherited copies of each chromosome into haplotypes and unlocks novel analyses at the haplotype level. In this work, we describe a new phasing method, SHAPEIT5, that accurately and rapidly phases large sequencing datasets and illustrates its key features on the UK Biobank WGS and WES data. First, we show that it phases rare variants with high accuracy. For instance, variants found in 1 sample out of 100,000 in the WES data are phased with accuracy above 95%. Second, we show that it can phase singletons, although with moderate accuracy, thereby making their inclusion in downstream analyses possible. Third, we show that the use of UK Biobank as a reference panel increases the accuracy of genotype imputation, an increase that is more pronounced when phased with SHAPEIT5 compared to other methods. Finally, we screen the phased WES data for loss-of-function (LoF) compound heterozygous (CH) events and identify 549 genes in which both gene copies are found knocked out. This list of genes complements current knowledge of gene essentiality in the human genome. We provide SHAPEIT5 in an open-source format, providing researchers with the means to leverage haplotype information in genetic studies.

genomics↗

bioRxiv2022

Widespread enhancer co-activity identified by multimodal single cell analysis

Non-coding regulatory elements such as enhancers are key in controlling the cell type-specificity and spatio-temporal expression of genes. To drive stable and precise gene transcription that is robust to genetic variation and environmental stress, genes are often targeted by multiple enhancers with redundant action. However, it is unknown whether enhancers targeting the same gene display simultaneous activity or whether some enhancer combinations are more often co-active than others. Here, we take advantage of the recent developments in single cell technology that permit assessing chromatin status (scATAC-seq) and gene expression (scRNA-seq) in the same single cells to link gene expression to the activity of multiple enhancers. Measuring activity patterns across 24,844 human lymphoblastoid single cells, we found that the majority of enhancers associated with the same gene display significant correlation in their chromatin profiles. For 6944 expressed genes associated with enhancers, we identified 89,885 significant enhancer-enhancer associations between nearby enhancers. We found that associated enhancers share similar transcription factor binding profiles and that gene essentiality is linked with higher enhancer co-activity. Our extensive enhancer co-activity maps can be used to pinpoint combinations of enhancers relevant in gene expression regulation and allow us to better predict the effect of genetic variation falling in non-coding regions.

genomics↗

bioRxiv2022

Genetic variation in correlated regulatory region of Immunity

Studying the interplay between genetic variation, epigenetic changes and regulation of gene expression in immune cells is important to understand the modification of cellular states in various conditions, including immune diseases. Here, we built cis maps of regulatory regions with coordinated activity - Cis Regulatory Domains (CRDs) - in neutrophils, monocytes and T cells. For this, we leveraged (i) whole-genome sequencing (WGS), (ii) chromatin immunoprecipitation sequencing (ChIP-seq), (iii) DNA methylation (450k arrays), and (iv) transcriptional profiles (RNA-seq) from the BLUEPRINT consortium, for up to 200 individuals. Our study uncovers 9287, 7666 and 5480 histone CRDs (hCRDs) and 6053, 6112, 5701 methyl CRDs (mCRDs) in monocytes, neutrophils and T-cells, respectively. We discovered 15294 hCRD-gene and 6185 mCRD-gene associations (5% FDR). Only 33% of hCRD-gene associations and 37% of mCRD-gene associations were shared between cell-types, revealing the dynamic nature of regulatory interactions and how similarly located regulatory regions modulate the activity of different genes on different cell types. We mapped Quantitative Trait Loci associated with CRD activity (CRD-QTLs) and found that 89% and 70% of these hCRDs and mCRDs are under genetic control highlighting the importance of genetic variation to study the coordination of cellular regulatory programs. We found CRD-QTLs to be enriched in celltype-specific transcription factor binding sites, such as SPI1, STAT3, RFX1, SOX4, ATF3 for neutrophils and monocytes and TCF4 and BCL11A for T-cells, in line with the Human protein Atlas. We integrated PCHi-C data, which showed that most significant associations discovered within gene-CRD associations and co-expressed genes associated with the same CRD, involving large genomic distances, tend to happen between genomic regions in close spatial proximity. Finally, we mapped trans regulatory associations between CRDs, which enabled the discovery of 207 trans-eQTLs across cell types. Overlapping our hits with trans eQTLs from eQTLGen Consortium meta-analysis in whole blood revealed 52 trans-eQTLs shared between the two studies. Overall, we show that mapping functional regulatory units using population genomics data allows discovering important mechanisms in the regulation of gene expression in immune cells and gain a greater understanding of cell-type specific regulatory mechanisms of immunity.

bioinformatics↗

bioRxiv2022

SELF PRUNING 3C is a flowering repressor that modulates seed germination, root architecture and drought responses

Allelic variation in the CETS (CENTRORADIALIS, TERMINAL FLOWER 1, SELF PRUNING) gene family has been shown to control agronomically important traits in many crops. CETS genes encode phosphatidylethanolamine binding proteins (PEBPs) that have a central role in flowering time control as florigenic and anti-florigenic signals. The great expansion of CETS genes in many species suggests that the functions of this family go beyond flowering. Here, we characterize the tomato SELF PRUNING 3C (SP3C) gene, and show that besides acting as a flowering repressor it also regulates seed germination and modulates root architecture. We show that loss of SP3C function in CRISPR/Cas9-generated mutant lines accelerates seed germination and increases root length with lower root side branching. Higher SP3C expression in transgenic lines promotes the opposite effects and also improves tolerance to water stress in seedlings. These discoveries provide insights into the role of SP paralogs in agronomically relevant traits and support future exploration of the involvement of CETS genes in abiotic stress responses. HighlightThe SELF PRUNING 3C (SP3C) gene is a repressor of flowering in tomato and exhibits additional functions, acting as a repressor of seed germination and modulating root architecture.

molecular biology↗

bioRxiv2021

Shared regulation and functional relevance of local gene co-expression revealed by single cell analysis

Most human genes are co-expressed with a nearby gene. Yet, previous studies only reported this extensive local gene co-expression using bulk RNA-seq. Here, we leverage single cell datasets in >85 individuals to identify gene co-expression across cells, unbiased by cell type heterogeneity and benefiting from the co-occurrence of transcription events in single cells. We discover thousands of co-expressed genes in two cell types and (i) compare single cell to bulk RNA-seq in identifying local gene co-expression, (ii) show that many co-expressed genes - but not the majority - are composed of functionally-related genes and (iii) provide evidence that these genes are transcribed synchronously and their co-expression is maintained up to the protein level. Finally, we identify gene-enhancer associations using multimodal single cell data, which reveal that >95% of co-expressed gene pairs share regulatory elements. Our in-depth view of local gene co-expression and regulatory element co-activity advances our understanding of the shared regulatory architecture between genes.

genomics↗

bioRxiv2021

Parent-of-origin effects in the UK Biobank

Identical genetic variations can have different phenotypic effects depending on their parent of origin (PofO). Yet, studies focussing on PofO effects have been largely limited in terms of sample size due to the need of parental genomes or known genealogies. Here, we used a novel probabilistic approach to infer PofO of individual alleles in the UK Biobank that does not require parental genomes nor prior knowledge of genealogy. Our model uses Identity-By-Descent (IBD) sharing with second- and third-degree relatives to assign alleles to parental groups and leverages chromosome X data in males to distinguish maternal from paternal groups. When combined with robust haplotype inference and haploid imputation, this allowed us to infer the PofO at 5.4 million variants genome-wide for 26,393 UK Biobank individuals. We used this large dataset to systematically screen 59 biomarkers and 38 anthropomorphic phenotypes for PofO effects and discovered 101 significant associations, demonstrating that this type of effects contributes to the genetics of complex traits. Notably, we retrieved well known PofO effects, such as the MEG3/DLK1 locus on platelet count, and we discovered many new ones at loci often unsuspected of being imprinted and, in some cases, previously thought to harbour additive associations.

genomics↗

bioRxiv2020

The molecular basis, genetic control and pleiotropic effects of local gene co-expression

Nearby genes are often expressed as a group. Yet, the prevalence, molecular mechanisms and genetic control of local gene co-expression are far from being understood. Here, by leveraging gene expression measurements across 49 human tissues and hundreds of individuals, we found that local gene co-expression occurs in 13% to 53% genes per tissue. By integrating various molecular assays (e.g. ChIP-seq and Hi-C), we estimated the ability of several mechanisms, such as enhancer-gene targeting, in distinguishing gene pairs that are co-expressed from those that are not. Notably, we identified 32,636 expression quantitative trait loci (eQTLs) which associate to co-expressed gene pairs and often overlap enhancer regions. Due to affecting several genes, these eQTLs are more often associated with multiple human traits than other eQTLs. Our extensive search for local gene co-expression opens the way to comprehend trait pleiotropy and comorbidity and provide functional interpretation of QTL and GWAS findings.

genomics↗

bioRxiv2019

The role of 3’UTR-protein complexes in the regulation of protein multifunctionality and subcellular localization

Multifunctional proteins often perform their different functions when localized in different subcellular compartments. However, the mechanisms leading to their localization are largely unknown. Recently, 3UTRs were found to regulate the cellular localization of newly synthesized proteins through the co-translational formation of 3UTR-protein complexes. Here, we investigate the formation of 3UTR-protein complexes involving multifunctional proteins by exploiting large-scale protein-protein and protein-RNA interaction networks. Focusing on 238 human extreme multifunctional (EMF) proteins, we predicted 1411 3UTR-protein complexes involving 128 EMF proteins and evaluated their role in regulating protein cellular localization and multifunctionality. Notably, we find that EMF proteins lacking localization addressing signals, yet present at both the nucleus and cell surface, often form 3UTR-protein complexes. In addition, they provide EMF proteins with the diversity of interaction partners necessary to their multifunctionality. Archetypal moonlighting proteins are also predicted to form 3UTR-protein complexes thereby reinforcing our findings. Finally, our results indicate that the formation of 3UTR-protein complex may be a common phenomenon in human cells, affecting up to 20% of the proteins in the human interactome.

systems biology↗

bioRxiv2019

Elevated CO2 induces age-dependent restoration of growth and metabolism in gibberellin-deficient plants

Main conclusion The effect of elevated [CO2] on the growth of tomato plants with reduced GA content is influenced by developmental stage. The increase of carbon dioxide (CO2) in the atmosphere during the last decades has aroused interest in the function of this gas in the growth and development of plants. Despite the known association between elevated CO2 concentration ([CO2]) and plant growth, its effects in association with gibberellin (GA), plant hormone that regulates de major aspects of plant growth, are still poorly understood. Therefore, we evaluated the effect of elevated [CO2] on growth and primary metabolism in tomato plants with drastic reduction in GA content (gib-1) at two different growth stages (21 and 35 days after germination, dag). Disruption on growth, photosynthetic parameters and primary metabolism were restored when gib-1 plants were transferred to elevated [CO2] at 21 dag. Elevated [CO2] also stimulated growth and photosynthetic parameters in Wild type (WT) plants at 21 dag, however, minor changes were observed in the level of primary metabolites. At 35 dag, elevated [CO2] did not stimulate growth in WT plants and gib-1 mutants showed their characteristic stunted growth phenotype.

plant biology↗

bioRxiv2017

Bundle sheath extensions in tomato affect leaf phenotypic plasticity in response to irradiance

Coordination between structural and physiological traits is key to plants responses to environmental fluctuations. In heterobaric leaves, bundle sheath extensions (BSEs) increase photosynthetic performance (light-saturated rates of photosynthesis, Amax) and water transport capacity (leaf hydraulic conductance, Kleaf). However, it is not clear how BSEs affect these and other leaf developmental and physiological parameters in response to environmental conditions. The obscuravenosa (obv) mutation, found in many commercial tomato varieties, leads to absence of BSEs. We examined structural and physiological traits of tomato heterobaric and homobaric (obv) near-isogenic lines (NILs) grown at two different irradiance levels. Kleaf, minor vein density and stomatal pore area index decreased with shading in heterobaric but not in homobaric leaves, which show similarly lower values in both conditions. Homobaric plants, on the other hand, showed increased Amax, leaf intercellular air spaces and mesophyll surface area exposed to intercellular airspace (Smes) in comparison with heterobaric plants when both were grown in the shade. BSEs further affected carbon isotope discrimination, a proxy for long-term water-use efficiency. BSEs confer plasticity in traits related to leaf structure and function in response to irradiance levels and might act as a hub integrating leaf structure, photosynthetic function and water supply and demand.\n\nSummary statementThe presence of bundle sheath extension (BSEs) defines leaves as heterobaric, as opposed to homobaric leaves that lack them. Multiple functions have been proposed for BSEs, but their impact on different environmental conditions is still unclear. Here, we compared a tomato (Solanum lycopersicum) homobaric mutant lacking BSEs with its corresponding heterobaric wild-type, grown under two irradiance conditions. We show that the presence of BSEs differentially alters various physiological and anatomical parameters in response to growth irradiance. We propose that BSEs could act as hubs coordinating leaf plasticity in response to environmental factors.\n\nArticle typeResearch article

plant biology↗