Browsing by Author "Yuseff, Maria-Isabel"

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    B Cells Adapt Their Nuclear Morphology to Organize the Immune Synapse and Facilitate Antigen Extraction
    (2022) Ulloa, Romina; Corrales, Oreste; Cabrera-Reyes, Fernanda; Jara-Wilde, Jorge; Saez, Juan Jose; Rivas, Christopher; Lagos, Jonathan; Haertel, Steffen; Quiroga, Clara; Yuseff, Maria-Isabel; Diaz-Munoz, Jheimmy
    Upon interaction with immobilized antigens, B cells form an immune synapse where actin remodeling and re-positioning of the microtubule-organizing center (MTOC) together with lysosomes can facilitate antigen extraction. B cells have restricted cytoplasmic space, mainly occupied by a large nucleus, yet the role of nuclear morphology in the formation of the immune synapse has not been addressed. Here we show that upon activation, B cells re-orientate and adapt the size of their nuclear groove facing the immune synapse, where the MTOC sits, and lysosomes accumulate. Silencing the nuclear envelope proteins Nesprin-1 and Sun-1 impairs nuclear reorientation towards the synapse and leads to defects in actin organization. Consequently, B cells are unable to internalize the BCR after antigen activation. Nesprin-1 and Sun-1-silenced B cells also fail to accumulate the tethering factor Exo70 at the center of the synaptic membrane and display defective lysosome positioning, impairing efficient antigen extraction at the immune synapse. Thus, changes in nuclear morphology and positioning emerge as critical regulatory steps to coordinate B cell activation.
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    Medium-throughput image-based phenotypic siRNA screen to unveil the molecular basis of B cell polarization
    (2023) Obino, Dorian; Maurin, Mathieu; Dingli, Florent; Loew, Damarys; Lescure, Aurianne; Terriac, Emmanuel; Goudot, Christel; Malbec, Odile; Lankar, Danielle; Yuseff, Maria-Isabel; Lennon-Dumenil, Ana-Maria; Moreau, Helene D.
    Cell polarity is an essential and highly conserved process governing cell function. Cell polarization is generally triggered by an external signal that induces the relocation of the centrosome, thus defining the polarity axis of the cell. Here, we took advantage of B cells as a model to study cell polarity and perform a medium-throughput siRNA-based imaging screen to identify new molecular regulators of polarization. We first identified candidates based on a quantitative proteomic analysis of proteins differentially associated with the centrosome of resting non-polarized and stimulated polarized B cells. We then targeted 233 candidates in a siRNA screen and identified hits regulating the polarization of the centrosome and/or lysosomes in B cells upon stimulation. Our dataset of proteomics, images, and polarity indexes provides a valuable source of information for a broad community of scientists interested in the molecular mechanisms regulating cell polarity.
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    SNX5 promotes antigen presentation in B cells by dual regulation of actin and lysosomal dynamics
    (2024) Cabrera-Reyes, Fernanda; Contreras-Palacios, Teemly; Ulloa, Romina; Jara-Wilde, Jorge; Caballero, Mia; Quiroga, Clara; Feijoo, Carmen G.; Diaz-Munoz, Jheimmy; Yuseff, Maria-Isabel
    B cells rapidly adapt their endocytic pathway to promote the uptake and processing of extracellular antigens recognized through the B-cell receptor (BCR). The mechanisms coupling changes in endomembrane trafficking to the capacity of B cells to screen for antigens within lymphoid tissues remain unaddressed. We investigated the role of SNX5, a member of the sorting nexin family, which interacts with endocytic membranes to regulate vesicular trafficking and macropinocytosis. Our results show that in steady state, B cells form SNX5-rich protrusions at the plasma membrane, which dissipate upon interaction with soluble antigens, whereas B cells activated with immobilized antigens accumulate SNX5 at the immune synapse where it regulates actin-dependent spreading responses. B cells silenced for SNX5 exhibit enlarged lysosomes, which are not recruited to the synaptic membrane, decreasing their capacity to extract immobilized antigens. Overall, our findings reveal that SNX5 is critical for actin-dependent plasma membrane remodeling in B cells involved in antigen screening and immune synapse formation, as well as endolysosomal trafficking required to promote antigen extraction and presentation.