Biology subjects

Duhamel, S.

Publications and source records attributed to Duhamel, S..

8 recordsLinked to original sources

bioRxiv2026

Compositional Heterogeneity Structures Microbial Microhabitats across Distinct Mineral Substrates

Microbial communities living on and in rocks operate at the microscale, where interactions with minerals fundamentally shape community structure and function. Yet the relationship between micron scale mineralogical configurations and microbial distributions remains poorly understood. We tested the hypothesis that microbial biomass spatially correlates with areas of heightened mineralogical heterogeneity by applying Raman microspectroscopy to rock samples from three geologically distinct substrates: authigenic carbonates from a marine methane seep, volcanic basalt from Iceland, and polymetallic nodules from the abyssal seafloor. Using spectral decomposition and multiple complementary metrics of compositional heterogeneity, we evaluated intra-pixel and inter-pixel heterogeneity patterns in relation to biomass distribution. Our analyses reveal three patterns across all sample types. 1) When spectra are deconstructed into their constituent components, biomass zones are disproportionately dominated by the biomass spectral component compared with primary mineral components in zones of different minerals. 2) Biomass spectra have more homogeneous compositional profiles than mineral spectra. 3) Biomass is surrounded by more heterogeneous microhabitats than mineral pixels. These findings demonstrate that biomass exerts a distinctive and consistent influence on Raman spectral signatures, both within and between pixels, in ways that mineral components do not. Our results establish generalizable principles linking microscale mineralogical properties to microbial biogeography; these properties could be used as a potential biosignature and may provide a standardized workflow applicable to diverse rock systems and astrobiological exploration strategies.

microbiology↗

bioRxiv2026

AMPK-dependent FLCN phosphorylation activates a survival response to hypoxia

Adaptation to low oxygen tensions is required to survive physiological and pathological stress conditions. We discovered an adenosine monophosphate-activated protein kinase (AMPK)-dependent, hypoxia-specific survival pathway through transcription factors (TFEB, TFE3). This pathway is distinct from nutrient sensing and independent of GAP Activity Towards Rags 1 (GATOR1). Mechanistically, AMPK phosphorylates the tumor suppressor folliculin (FLCN) at a highly conserved serine, inhibiting its GAP activity towards the Rag-GTPases, resulting in mechanistic target of rapamycin complex 1 (mTORC1) inhibition, TFE3 activation, resulting in lysosomal and mitochondrial biogenesis essential for hypoxia adaptation. TFEB/3 activity correlates with the hypoxia signature in hypoxic tumor areas. Cancer patients with elevated TFEB/3 activity show poor survival, specifically in hypoxia. These findings identify the AMPK-FLCN-TFE3 axis as a therapeutic target in tumors.

biochemistry↗

bioRxiv2025

Divergent roles of DNA methylation, TRIM28, and p53 surveillance in human embryonic and trophoblast stem cells

Transcriptional regulation of transposons and genes by TRIM28 and 5mC is critical for proper mammalian embryonic development, but the specific roles for these mediators in human embryonic and placental lineage remain unclear. We find that loss of TRIM28 has a limited effect on global transposon expression and instead results in upregulation of genes proximal to TRIM28-bound Long Terminal Repeats (LTRs) in both human trophoblast stem cells (hTSCs) and human embryonic stem cells (hESCs). MER11A elements show especially strong regulatory importance in hTSCs: these elements are bound by both TRIM28 and placental transcription factors and show both heterochromatic and euchromatic features. Some genes are positively regulated by MER11A elements in hTSC basal state, while other MER11A-proximal genes show upregulation only upon TRIM28 deletion. By contrast, loss of DNA methylation in hESCs or hTSCs leads to a global increase in transposon expression. While many genic 5mC targets are shared in hESCs and hTSCs, we also observe evidence that a handful of genes important for somatic development are repressed by 5mC in trophoblast, while a small parallel set of placental genes are repressed by methylation in embryonic tissue. Interestingly, loss of DNMT1 causes hESCs to be rapidly lost from culture in a TP53 and mitotic surveillance checkpoint-dependent manner, while hTSCs show little p53 response to DNMT1 loss or DNA damage generally, instead showing gradual mitotic defects and aneuploidy and slow loss from culture. This discrepancy may explain the higher frequency of karyotypic abnormality found in human placental cells. Together, this study charts the role of TRIM28 and DNA methylation in regulating embryonic and placental transcription and demonstrates divergent p53-dependent responses to genomic instability.

molecular biology↗

bioRxiv2024

Dual-BONCAT Reveals Distinct Day- and Night-Active Microbial Populations in Salt Marsh Sediments

Bio-Orthogonal Non-Canonical Amino Acid Tagging (BONCAT) has emerged as a prominent molecular technique that enables microbial ecologists to visualize and identify metabolically active cells in cultures and complex microbial communities. To date, researchers have used just one non-canonical amino acid (ncAA) in a given experiment; here, we validate a novel approach using two different ncAAs in a single experiment. This advancement facilitates the detection of differentially active subpopulations within the same experimental context, thereby reducing the uncertainty and variability associated with parallel treatments, and providing precise spatial information about organisms that are active under distinct conditions or at different times. We show that both ncAAs can be taken up by E. coli cultures and by constituents of the Little Sippewissett Salt Marsh microbiome, resulting in fluorescence signals that are significantly higher than background and ncAA-free control experiments, as well as differential labeling patterns reflective of distinct subpopulations. As a proof of concept, we implemented this "dual-BONCAT" approach in salt marsh sediments, adding one ncAA during daytime hours and the other at night. Subpopulations of cells that were anabolically active during the day and/or night were distinguishable by both fluorescence microscopy, and by fluorescence-activated cell sorting. Subsequent high-throughput 16S rRNA gene amplicon sequencing of active subpopulations revealed that Methylobacterium, potentially feeding on plant exudate carbon, was preferentially active during the day, while sulfur-cycling taxa dominated the night-active population. Dual-BONCAT offers an important advancement in multiplexing substrate-analog probing techniques, providing a more realistic understanding of metabolic activity under distinct environmental conditions. ImportanceMicrobial communities are complex and dynamic, with different groups of microbes active under distinct conditions. Bio-Orthogonal Non-Canonical Amino Acid Tagging (BONCAT) uses synthetic amino acids to tag newly made proteins, allowing researchers to see and identify the active subset of a community. While BONCAT studies to date have used a single synthetic amino acid to evaluate cell activity in a single experimental context, here we introduce a new approach, "dual-BONCAT," using two synthetic amino acids to track differential responses to changing conditions. After validating the approach with E. coli, we deployed it in a salt marsh sediment community, finding that organisms potentially feeding on plant root sugars were more active during the day, and microbes likely metabolizing sulfur were more active at night. We believe dual-BONCAT will prove useful in many studies, as it illuminates microbial community responses to changing conditions, which has important implications for ecosystem dynamics.

microbiology↗

bioRxiv2021

Differential timing for glucose assimilation in Prochlorococcus and coexistent microbial populations at the North Pacific Subtropical Gyre

The marine cyanobacterium Prochlorococcus can utilize glucose as a source of carbon. However, the relative importance of inorganic and organic carbon assimilation and the timing of glucose assimilation are still poorly understood in these numerically dominant cyanobacteria. Here we investigated whole microbial community and group-specific primary production and glucose assimilation, using incubations with radioisotopes combined with flow cytometry cell sorting. We also studied changes in the microbial community structure in response to glucose enrichments and analyzed the transcription of Prochlorocccus genes involved in carbon metabolism and photosynthesis. Our results showed a circadian rhythm for glucose assimilation in Prochlorococcus, with maximum assimilation during the midday and minimum at midnight, which was different compared with that of the total microbial community. This suggests that rhythms in glucose assimilation have been adapted in Prochlorococcus to couple the active transport to photosynthetic light reactions producing energy, and possibly to avoid competition from the rest of the microbial community. High-light Prochlorococcus strains showed most transcriptional changes upon glucose enrichment. Pathways involved in glucose metabolism as the pentose phosphate, the Entner-Dudoroff, glycolysis, respiration and glucose transport showed an increase in the transcript level. A few genes of the low-light strains showed opposite changes, suggesting that glucose assimilation has been subjected to diversification along the Prochlorococcus evolution.

microbiology↗

bioRxiv2021

Met-HER3 crosstalk supports proliferation via MPZL3 in MET-amplified cancer cells

Receptor tyrosine kinases (RTKs) are recognized as targets of precision medicine in human cancer upon their gene amplification or constitutive activation, resulting in increased downstream signal complexity including heterotypic crosstalk with other RTKs. The Met RTK exhibits such reciprocal crosstalk with several members of the human EGFR (HER) family of RTKs when amplified in cancer cells. We show that Met signaling converges on HER3 tyrosine phosphorylation across a panel of seven MET-amplified cancer cell lines and that HER3 is required for cancer cell expansion and oncogenic capacity in-vitro and in-vivo. Gene expression analysis of HER3-depleted cells identified MPZL3, encoding a single-pass transmembrane protein, as a HER3-dependent effector in multiple MET-amplified cancer cell lines. MPZL3 interacts with HER3 and MPZL3 loss phenocopies HER3 loss in MET-amplified cells, while MPZL3 overexpression rescues proliferation upon HER3 depletion. Together, these data support an oncogenic role for a HER3-MPZL3 axis in MET-amplified cancers.

cancer biology↗

bioRxiv2020

Changes in salt-marsh vegetation weakly affect top consumers of aquatic food webs

Salt marshes are under high, and increasing, anthropogenic pressures that have notably been reported to affect the diet of several fish species, probably resulting in nursery function alterations. Most of the previous studies in Europe were yet based on gut content analysis of fish, which can be considered a snapshot of immediate impacts of salt-marsh changes, and hardly of long-term effects of disturbances. In this study, we investigated the impact of vegetation type (resulting from both plant invasion and sheep grazing) by assessing trophic network (and especially fish diet and position) of different salt-marsh conditions. Replicated samples of basic sources (particular organic matter and microphytobenthos), dominant vegetation, potential aquatic and terrestrial prey and fish of 3 main species were taken during summer 2010 in two bays from Western France (Mont -Saint-Michel Bay and Seine Estuary) and analysed using C and N stable isotope compositions. All response variables tested (overall trophic organization, trophic niche and trophic position) provided consistent results, i.e. a dominant site effect and a weaker effect of vegetation type. Site effect was attributed to differences in anthropogenic Nitrogen inputs and tidal regime between the two bays, with more marine signatures associated with a higher frequency of flooding events. A second hypothesis is that E. acuta, which has recently totally replaced typical salt-marsh vegetation in Mont Saint-Michel Bay strongly impacted the nursery function. The trophic status of dominant fish species was unchanged by local salt-marsh vegetation, and considered consistent with their diet, i.e. high for predatory species (the sea bass Dicentrarchus labrax and the common goby Pomatoschistus microps) and lower for biofilm grazing species (the thinlip mullet Chelon ramada). This study finally highlights the relevance of stable isotopes analyses for assessing long-term and integrative effects of changes in vegetation resulting from human disturbances in salt marshes. HighlightsO_LICross-ecosystem subsidies are of high functional importance, notably in salt marshes C_LIO_LIFish are vectors of exchanges, most European studies being based on their gut content C_LIO_LIUsing stable isotopes we analysed the effect of surrounding vegetation on food webs C_LIO_LISurprisingly we found weak vegetation and strong site effects on all metrics C_LIO_LINitrogen inputs, site accessibility and loss of nursery function can explain this fact C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=87 SRC="FIGDIR/small/222406v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@27b54borg.highwire.dtl.DTLVardef@ef583corg.highwire.dtl.DTLVardef@18a0303org.highwire.dtl.DTLVardef@1cbe5d4_HPS_FORMAT_FIGEXP M_FIG C_FIG

ecology↗

bioRxiv2020

Heterotrophic Carbon Fixation in a Salamander-Alga Symbiosis.

The unique symbiosis between a vertebrate salamander, Ambystoma maculatum, and unicellular green alga, Oophila amblystomatis, involves multiple modes of interaction. These include an ectosymbiotic interaction where the alga colonizes the egg capsule, and an intracellular interaction where the alga enters tissues and cells of the salamander. One common interaction in mutualist photosymbioses is the transfer of photosynthate from the algal symbiont to the host animal. In the A. maculatum-O. amblystomatis interaction, there is conflicting evidence regarding whether the algae in the egg capsule transfer chemical energy captured during photosynthesis to the developing salamander embryo. In experiments where we took care to separate the carbon fixation contributions of the salamander embryo and algal symbionts, we show that inorganic carbon fixed by A. maculatum embryos reaches 2% of the inorganic carbon fixed by O. amblystomatis algae within an egg capsule after 2 hours in the light. After 2 hours in the dark, inorganic carbon fixed by A. maculatum embryos is 800% of the carbon fixed by O. amblystomatis algae within an egg capsule. Using photosynthesis inhibitors we show that A. maculatum embryos and O. amblystomatis algae compete for available inorganic carbon within the egg capsule environment. Our results confirm earlier studies suggesting a role of heterotrophic carbon fixation during vertebrate embryonic development. Our results also show that the considerable capacity of developing A. maculatum embryos for inorganic carbon fixation precludes our ability to distinguish any minor role of photosynthetically transferred carbon from algal symbionts to host salamanders using bicarbonate introduced to the egg system as a marker.

ecology↗