Biology subjects

Joly, N.

Publications and source records attributed to Joly, N..

6 recordsLinked to original sources

bioRxiv2026

The Endoplasmic Reticulum as a Cytoplasmic Anchor for Microtubule-Driven Aster Centration

The proper positioning of microtubule (MT) asters is essential to regulate division positioning and multi-cellular morphogenesis, yet the mechanisms regulating this process remain poorly understood. Using sea urchin zygotes as a model, we combined high-resolution imaging, perturbation assays, and 3D modeling to investigate the nature of forces driving sperm aster centration after fertilization. We show that centering asters are highly anisotropic, featuring long microtubules extending at the aster front that may engage with organelles in bulk cytoplasm to generate pulling forces. While vesicular compartments, like acidic organelles, mitochondria or yolk, contributed minimally, we found that the endoplasmic reticulum (ER) undergoes a massive compaction around the centering aster and drags cytosol to produce a reactive centering force on the aster. Perturbing ER organization by cytoplasmic stratification or Atlastin-mediated fragmentation disrupted aster centration. A two-fluid model suggests that the frictional interaction between the compacting ER and cytosol fluid is sufficient to recapitulate key features of aster centration. These findings identify the ER as an important cytoplasmic anchor for MT force exertion in bulk cytoplasm, to guide aster centering motion and influence early embryonic division geometries. More broadly, our results highlight the role of cytoplasmic architecture in shaping the mechanics of organelle positioning during development.

cell biology↗

bioRxiv2025

Molecular basis for the activation of Aurora A and Plk1 kinases during mitotic entry

The evolutionarily conserved, intrinsically disordered protein Bora is critical for initiating the activation of mitotic kinases. Once phosphorylated at Ser112 by Cyclin A-Cdk1 kinase, phospho-(p)Bora activates unphosphorylated Aurora A kinase (AURKA), directing it towards Polo-like kinase 1 (Plk1), thus promoting Cyclin B-Cdk1 activation and mitotic entry. Here, by combining structural modelling and in vitro assays, we provide evidence that Bora wraps around the N-terminal lobe of AURKA to position its phospho-Ser112 near AURKAs T-loop, mimicking T-loop phosphorylation. Additionally, Bora transiently interacts with the Alpha C helix of the Plk1 kinase domain through a conserved motif, guiding AURKA activity towards the Plk1 T-loop, which is otherwise impervious to phosphorylation by AURKA. We highlight the importance of this motif for Bora function in vitro and during mitotic entry in Xenopus egg extracts. Our results reveal critical molecular details of mitotic kinase activation, which could lead to the development of pathway-specific inhibitors. HighlightsO_LIMITOKINAC: a versatile approach that reconstitutes Bora and AURKA-dependent Plk1 T-loop phosphorylation in E. coli. C_LIO_LIAlphaFold 3 structural modeling of the phospho-Bora - AURKA complex tested by MITOKINAC using 39 Bora variants. C_LIO_LIPhospho-Bora wraps around the N-terminal lobe of AURKA to position its essential phospho-Serine 112 in the activation loop of AURKA. C_LIO_LIpBora delivers the AURKA activity to the Plk1 T-loop, which is otherwise impervious to AURKA phosphorylation. C_LI

biochemistry↗

bioRxiv2024

Direct observation of fluorescent proteins in gels: a rapid, cost-efficient and quantitative alternative to immunoblotting

The discovery of Green Fluorescent Protein (GFP) and its derivatives has revolutionized cell biology. These fluorescent proteins (FPs) have enabled the real-time observation of protein localization and dynamics within live cells. Applications of FP vary from monitoring gene/protein expression patterns, visualizing protein-protein interactions, measuring protein stability, assessing protein mobility and creating biosensors. The utility of FPs also extends to biochemical approaches through immunoblotting and proteomic analyses, aided by anti-FP antibodies and nanobodies. FPs are notoriously robust proteins with a tightly folded domain that confers a strong stability and a relative resistance to degradation and denaturation. In this study, we report that various green, orange and red FPs can be maintained in a native, fluorescent state during the entire process of protein sample extraction, incubation with sample buffer, loading and migration on SDS-PAGE with only minor adaptations of traditional protocols. This protocol results in the ability to detect and quantify in-gel fluorescence (IGF) of endogenously-expressed proteins tagged with FPs directly after migration, using standard fluorescence-imaging devices. This approach eliminates the need for antibodies and chemiluminescent reagents, as well as the time-consuming steps inherent in immunoblotting such as transfer onto a membrane and antibody incubations. Overall, IGF detection provides clearer data with less background interference, a sensitivity comparable or better to antibody-based detection, a better quantification and a broader dynamic range. After fluorescence imaging, gels can still be used for other applications such as total protein staining or immunoblotting if needed. It also expands possibilities by allowing the detection of FPs for which antibodies are not available. Our study explores the feasibility, limitations, and applications of IGF for detecting endogenously expressed proteins in cell extracts, providing insights into sample preparation, imaging conditions, and sensitivity optimizations, and potential applications such as co-immunoprecipitation experiments.

biochemistry↗

bioRxiv2023

Decoding Ten Years of Little Penguin Foraging: Bio-Logging Reveals Foraging Patterns with Implications for Climate Change Mitigation and Marine Spatial Planning

O_LIProtected areas are a widely adopted resource management strategy for mitigating the consequences of global change and preserve functioning ecosystems. Long-term species monitoring programmes, aided by bio-logging technology, provide insights into the extent and spatial variation of areas occupied by wild animals and inform conservation and management. High-resolution GPS-acceleration data offer a more accurate understanding of animal behavior and area use, compared to location-based inference, emphasizing the significance of specific sites amid long-term climate change. C_LIO_LIWe based our case-study on the largest colony of little penguins (Eudyptula minor) located at Phillip Island. Based on a ten-year bio-logging dataset (247 individual tracks), we combine high-resolution bio-logging data from GPS-accelerometer loggers with proxies for resource availability (e.g. Sea Surface Tenperature, thermocline, water turbidity). Using machine learning techniques and Generalized Additive Mixed Models, we quantify the environmental factors determining spatio-temporal variability in foraging effort (defined as hunting time) across different breeding seasons and stages. C_LIO_LILittle penguins increased their hunting time by reducing spatial displacement (shorter step length) and diving deeper, with a slower increase in hunting effort below 10 m depth. In relation to environmental conditions, penguins increased hunting effort in coastal areas with high turbid and productive waters and decreased effort with increasing Sea Surface Temperature. This gives insights into how these animals allocate effort differently according to shifting environmental conditions. C_LIO_LIOur analysis offers crucial long-term insights into little penguin area usage in the Bass Strait at sufficient spatial and temporal resolution for management and conservation planning. The Bass Strait is facing intense climatic and anthripogenic pressures, and the findings here on intensity of area usage and strategy shifting according to environmental conditions, are of great relevance for the marine spatial planning currently under development along the coast. C_LIO_LIPolicy implications: High-resolution behavioral information obtained from bio-logging data using GPS-accelerometer tags provides understanding of how species shift strategies in response to environmental variability. This is vital to implement climate-adaptive conservation and management strategies. Given the growing availability of long-term accelerometer datasets within the ecological community, we recommend integrating such high-resolution information into conservation programs. C_LI

ecology↗

bioRxiv2023

COVID-related anthropause highlights the impact of marine traffic on breeding little penguins

The COVID-19 pandemic and its lock-down measures have resulted in periods of reduced human activity, known as anthropause. While this period was expected to be favorable for the marine ecosystem, due to a probable reduction of pollution, shipping traffic, industrial activity and fishing pressure, negative counterparts such as the increased use of disposable plastic and reduced fisheries surveillance and enforcement could counterbalance these positive effects. Simultaneously, on-land pressure due to human disturbance and tourism should have drastically decreased, potentially benefiting land-based marine breeders such as seabirds. Thus, long-term datasets became crucial to differentiate between historical trends and any evident changes resulting from the anthropause. We analyzed 11 years of data on several biological parameters of little penguins (Eudyptula minor) from the Penguin Parade (R), a popular tourist attraction at Phillip Island, Australia. We investigated the impact of anthropogenic activities on penguin behavior during the breeding season measured by (1) distribution at sea, (2) colony attendance, (3) isotopic niche (4) chick meal mass, and (5) offspring investment against shipping traffic and number of tourists. The 2020 lock-downs resulted in a near absence of tourists visiting the Penguin Parade (R), which was otherwise visited by 800,000+ visitors on average per year. However, our long-term analysis showed no effect of the presence of visitors on little penguins activities. Surprisingly, the anthropause did not triggered any changes in maritime traffic intensity and distribution in the region. While we found significant inter- and intra-annual variations for most parameters, we detected a negative effect of marine traffic on the foraging efficiency. Our results suggest that environmental variations have a greater influence on the breeding behavior of little penguins compared to short-term anthropause events. Our long-term dataset was key to test whether changes in anthropogenic activities affected the wildlife during the COVID-19 pandemic. Impact statementWe found that marine traffic, but not tourist presence, negatively impact the foraging and provisioning behavior of little penguins.

ecology↗

bioRxiv2021

Cortical microtubule pulling forces contribute to the union of the parental genomes in the C. elegans zygote

Previously, we reported that the Polo-like kinase PLK-1 phosphorylates the single C. elegans lamin (LMN-1) to trigger lamina depolymerization during mitosis. We showed that this event is required for the formation of a pronuclear envelopes scission event that removes membranes on the juxtaposed oocyte and sperm pronuclear envelopes in the zygote, allowing the parental chromosomes to merge in a single nucleus after segregation (Velez-Aguilera, 2020). Here we show that cortical microtubule pulling forces contribute to pronuclear envelopes scission by promoting mitotic spindle elongation. We also demonstrate that weakening of the pronuclear envelopes, via PLK-1-mediated lamina depolymerization, is a prerequisite for the astral microtubule pulling forces to trigger pronuclear membranes scission. Finally, we provide evidence that PLK-1 mainly acts via lamina depolymerization in this process. These observations thus indicate that temporal coordination between lamina depolymerization and mitotic spindle elongation facilitates pronuclear envelopes scission and parental genomes unification.

cell biology↗