Biology subjects

Perez-Berna, A. J.

Publications and source records attributed to Perez-Berna, A. J..

5 recordsLinked to original sources

bioRxiv2026

A single Omicron mutation reshapes ORF3a-driven host-cell remodelling

SARS-CoV-2 ORF3a remodels host membranes, but the structural basis and metabolic consequences of this process remain unclear. Here, we combine complementary imaging approaches to define ORF3a function at nanometric scale, identifying underlying mechanisms, and determining how Omicron variant rewire this activity. ORF3a from the ancestral Wuhan strain disrupts Golgi cisternae, drives the formation of ORF3a dense vesicles, remodels mitochondrial architecture, and promotes lipid droplet expansion. Multi-omics analyses further reveal selective triacylglycerol accumulation linked to DGAT1 upregulation, which we validate pharmacologically through DGAT1 inhibition. In contrast, Omicron ORF3a variant, despite carrying only the Thr223Ile substitution within the {beta}7-{beta}8 loop at the bottom of the cytosolic domain, induced a dramatic phenotypic shift: ORF3a localizes to multivesicular bodies, preserves Golgi architecture, and fails to induce lipid accumulation. All together, these results identify ORF3a as a regulator of membrane organization and lipid homeostasis, showing how minimal sequence variation rewires host-cell remodelling. Graphical TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/742305v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@135032borg.highwire.dtl.DTLVardef@1633d4borg.highwire.dtl.DTLVardef@4d4115org.highwire.dtl.DTLVardef@1ebdf65_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2026

Correlative Synchrotron X-ray Microscopy Reveals Dose- and Division-Dependent Nanoparticle Redistribution in Macrophages

Understanding the intracellular fate of nanoparticles is essential for designing safer and more effective nanomedicines, yet most studies rely on static observations and lack high-resolution, near-native volumetric information. Here, we establish a synchrotron-based correlative X-ray microscopy framework to investigate how fluorescent silica nanoparticles (SiNPs) redistribute within macrophages as a function of concentration and successive cell-division cycles. SiNPs were internalized by RAW 264.7 macrophages at different concentrations and analyzed using a synchrotron-based correlative X-ray microscopy workflow integrating cryogenic soft X-ray tomography (cryo-SXT), cryogenic structured illumination microscopy (cryo-SIM), and coherent X-ray ptychography, with confocal fluorescence microscopy used to establish population-level uptake tendencies. Cryo-SXT reveals a concentration-dependent redistribution of nanoparticle-containing vesicles from peripheral endosomes toward the perinuclear region, while correlative cryo-SIM confirms strict vesicular confinement, with no evidence of free nanoparticle diffusion into the nucleoplasm. At higher doses, nanoparticles approach the nuclear region via vesicles extending into nuclear-envelope invaginations, rather than by true nuclear entry. Successive cell divisions redistribute the intracellular nanoparticle load and promote stable perinuclear clustering, identifying a long-term sequestration route in macrophages. Coherent X-ray ptychography further reveals nanoscale deformations of the nuclear envelope associated with dense perinuclear vesicles. Together, these results establish synchrotron-based correlative X-ray microscopy as a mechanistic, multiscale platform for unveiling the dynamic intracellular fate of nanoparticles and providing mechanistic insight into their apparent nuclear localization.

cell biology↗

bioRxiv2026

PAK-family kinases promote cell fusion irreversibility by preventing cell wall repair

Cell-cell fusion is critical for the sexual life cycle, as it drives the unidirectional transition between haploid and diploid phases. In fungi, fusion requires not only plasma membrane merging but also local cell wall removal. However, the integrity of the cell wall, which resists internal turgor pressure and is essential for survival, is normally monitored by a cell wall integrity pathway (CIP) involving Rho1 GTPase, protein kinase C (PKC) and MAPK signaling, which promotes damage repair. How the CIP allows localized cell wall degradation is not known. In the fission yeast Schizosaccharomyces pombe, we previously identified pak2{Delta} mutants that exhibit transient fusion: cells briefly fuse and exchange cytoplasmic contents, which signal catastrophic haploid meiosis, but then reseal their fusion pore. Here, we show that PAK-family kinase activity is essential for cell-cell fusion, with major and minor contribution for Pak2 and the essential Pak1, respectively. Strikingly, osmotic stabilization largely suppresses all PAK-associated fusion defects, pointing to a role in cell-wall remodeling. Pak2 accumulates at the fusion site, where it promotes fusion pore opening and expansion. In its absence, Rho1-GTP, PKC and beta-glucan synthase Bgs4 show elevated levels at the fusion site, and their partial loss-of-function rescues fusion. Pak1 mutants have elevated MAPK activity. Thus, Pak2 and Pak1 both antagonize the CIP. Furthermore, correlative light and cryo-soft-X-ray tomography reveals that resealed pores in PAK mutants rebuilt an intact cell wall. We conclude that PAKs enforce fusion directionality by antagonizing cell-wall repair mechanisms that otherwise restore separation between mating partners.

cell biology↗

bioRxiv2025

Identifying the internalization pathways of magnetotactic bacteria and magnetosomes by cancer cells

Given the need for new tumor treatment strategies, therapies using magnetic nanoparticles and bacteria are gaining momentum. In this context, magnetotactic bacteria and magnetosomes could act as theranostic agents for use in magnetic hyperthermia, targeted drug delivery, and magnetic resonance imaging. Thus, understanding their interaction with target cells is essential to ensure their theranostic efficiency. This study investigates the uptake of magnetotactic bacteria (MSR-1) and magnetosomes by lung carcinoma cells (A549). First, MSR-1 and magnetosomes are imaged inside A549 cells using cryo soft X-ray tomography, revealing the presence of MSR-1 and magnetosomes inside endosomes. Subsequently, the endocytosis pathways involved in the internalization of MSR-1 and magnetosomes by the cells are elucidated. It is observed that MSR-1 mainly enter cells by receptor-mediated endocytosis, as described previously for other intracellular bacteria. However, the endocytosis of magnetosomes occurs mainly via phagocytosis or macropinocytosis, probably due to the large size of the formed magnetosome clusters. These findings fill a key gap in our understanding of the internalization of MSR-1 bacteria and magnetosomes by lung carcinoma cells, and establish a method applicable to studying their internalization by other target cell types. Graphical AbstractMagnetotactic bacteria (Magnetospirillum gryphiswaldense MSR-1) and magnetosomes, which are proposed as cancer theranostic mediators, are imaged inside A549 cancer cells in 3D using cryo soft X-ray tomography. Moreover, the endocytosis pathways that the cells use to internalize MSR-1 and magnetosomes are elucidated. MSR-1 predominantly enter A549 cells via receptor-mediated endocytosis, while magnetosomes are preferentially internalized via receptor-independent endocytosis. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=102 SRC="FIGDIR/small/690460v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@bbcac0org.highwire.dtl.DTLVardef@fc33d6org.highwire.dtl.DTLVardef@1af65aeorg.highwire.dtl.DTLVardef@3ca539_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2020

Four-dimensional characterization of the Babesia divergens asexual life cycle: from the trophozoite to the multiparasite stage

Babesia is an apicomplexan parasite of significance that causes the disease known as babesiosis in domestic and wild animals and in humans worldwide. Babesia infects vertebrate hosts and reproduces asexually by a form of binary fission within erythrocytes/red blood cells (RBCs), yielding a complex pleomorphic population of intraerythrocytic parasites. Seven of them, clearly visible in human RBCs infected with Babesia divergens, are considered the main forms and named single, double and quadruple trophozoites, paired and double paired-pyriforms, tetrad or Maltese Cross, and multiparasite stage. However, these main intraerythrocytic forms coexist with RBCs infected with transient parasite combinations of unclear origin and development. In fact, little is understood about how Babesia builds this complex population during its asexual life cycle. By combining the emerging technique cryo soft X-ray tomography and video microscopy, main and transitory parasites were characterized in a native whole cellular context and at nanometric resolution. As a result, the architecture and kinetic of the parasite population has been elucidated. Importantly, the process of multiplication by binary fission, involving budding, was visualized in live parasites for the first time, revealing that fundamental changes in cell shape and continuous rounds of multiplication occur as the parasites go through their asexual multiplication cycle. Based on these observations, a four-dimensional (4D) asexual life cycle model has been designed highlighting the origin of the tetrad, double paired-pyriform and multiparasite stages and the transient morphological forms that, surprisingly, intersperse in a chronological order between one main stage and the next along the cycle. IMPORTANCEBabesiosis is a disease caused by intraerythrocytic Babesia parasites, which possess many clinical features that are similar to those of malaria. This worldwide disease, is increasing in frequency and geographical range, and has a significant impact on human and animal health. Babesia divergens is one of the species responsible for human and cattle babesiosis causing death unless treated promptly. When B. divergens infects its vertebrate hosts it reproduces asexually within red blood cells. During its asexual life cycle, B. divergens builds a population of numerous intraerythrocytic (IE) parasites of difficult interpretation. This complex population is largely unexplored, and we have therefore combined three and four dimensional (3D and 4D) imaging techniques to elucidate the origin, architecture, and kinetic of IE parasites. Unveil the nature of these parasites have provided a vision of the B. divergens asexual cycle in unprecedented detail and a key step to develop control strategies against babesiosis

microbiology↗