A5024938116- Cellular transport and secretion
- Endoplasmic Reticulum Stress and Disease
- Lipid Membrane Structure and Behavior
- Lysosomal Storage Disorders Research
- Trypanosoma species research and implications
- Glycosylation and Glycoproteins Research
- Sphingolipid Metabolism and Signaling
- Calcium signaling and nucleotide metabolism
- Autophagy in Disease and Therapy
- Microbial Metabolic Engineering and Bioproduction
- RNA and protein synthesis mechanisms
- Lipid metabolism and biosynthesis
- RNA regulation and disease
- Biofuel production and bioconversion
- Plant Virus Research Studies
- GABA and Rice Research
- Metabolomics and Mass Spectrometry Studies
- Social Skills and Education
- Educational Research and Science Teaching
- CRISPR and Genetic Engineering
- Educational Innovations and Technology
- Peptidase Inhibition and Analysis
Universidad de Sevilla
2008-2023
Hospital Universitario Virgen del Rocío
2020-2023
Instituto de Biomedicina de Sevilla
2020-2023
Hiroshima University
2021
NCCR Chemical Biology - Visualisation and Control of Biological Processes Using Chemistry
2014-2020
University of Geneva
2013-2020
European Molecular Biology Laboratory
2015
To elucidate new functions of sphingosine (Sph), we demonstrate that the spontaneous elevation intracellular Sph levels via caged leads to a significant and transient calcium release from acidic stores is independent 1-phosphate, extracellular ER levels. This photo-induced Sph-driven requires two-pore channel 1 (TPC1) residing on endosomes lysosomes. Further, uncaging translocation autophagy-relevant transcription factor EB (TFEB) nucleus specifically after lysosomal release. We confirm...
Glycosylphosphatidylinositol (GPI)-anchored proteins are secretory that attached to the cell surface of eukaryotic cells by a glycolipid moiety. Once GPI anchoring has occurred in lumen endoplasmic reticulum (ER), structure lipid part on anchor undergoes remodeling process prior ER exit. In this study, we provide evidence suggesting yeast p24 complex, through binding specifically GPI-anchored an anchor-dependent manner, plays dual role their selective trafficking. First, complex promotes...
The regulatory pathways required to maintain eukaryotic lipid homeostasis are largely unknown. We developed a systematic approach uncover new players in the regulation of homeostasis. Through an unbiased mass spectrometry-based lipidomic screening, we quantified hundreds species, including glycerophospholipids, sphingolipids, and sterols, from collection 129 mutants protein kinase phosphatase genes Saccharomyces cerevisiae. Our successfully identified known kinases involved uncovered ones....
3D high-resolution live imaging reveals the importance of ceramide chain length for protein sorting in selective export sites.
Lag1 was the first longevity assurance gene, discovered in Saccharomyces cerevisiae. is a ceramide synthase and its homolog, Lac1, has similar enzymatic function but no role aging. Lac1 lay an branchpoint of sphingolipid pathway that interconnected by activity C4-hydroxylase, Sur2. By uncoupling branch point using lipidomic mass spectrometry, metabolic labeling vitro assays we show preferentially synthesizes phyto-sphingolipids. Using photo-bleaching experiments protein uniquely required for...
The p24 family members are transmembrane proteins assembled into heteromeric complexes that continuously cycle between the ER and Golgi apparatus. These cargo were assumed to play a structural role in COPI budding because of their major presence mammalian vesicles. However, this putative function has not been proved conclusively so far. Furthermore, deletion all eight yeast does produce severe transport phenotypes, suggesting complex is essential for function. In paper we provide direct...
Endoplasmic reticulum (ER) quality control mechanisms target terminally misfolded proteins for ER-associated degradation (ERAD). Misfolded glycophosphatidylinositol-anchored (GPI-APs) are, however, generally poor ERAD substrates and are targeted mainly to the vacuole/lysosome degradation, leading predictions that a GPI anchor sterically obstructs ERAD. Here we analyzed of GPI-AP Gas1* in yeast. We could efficiently route Hrd1-dependent provide evidence it contains anchor, ruling out Instead,...
Lipids synthesized at the endoplasmic reticulum (ER) are delivered to Golgi by vesicular and non-vesicular pathways. ER-to-Golgi transport is critical for maintaining different membrane lipid composition identities of organelles. Despite their importance, mechanisms regulating remain elusive. Here we report that coat protein complex II (COPII) vesicle-mediated ceramide from ER requires yeast oxysterol-binding homologs, Osh proteins, which have been implicated in homeostasis. Because proteins...
Glycosylphosphatidylinositol-anchored proteins (GPI-APs) exit the endoplasmic reticulum (ER) through a specialized export pathway in yeast Saccharomyces cerevisiae. We have recently shown that very-long acyl chain (C26) ceramide present ER membrane drives clustering and sorting of GPI-APs into selective sites (ERES). Now, we show this lipid-based also involves C26 as lipid moiety GPI-APs, which is incorporated GPI anchor lipid-remodeling process after protein attachment ER. Moreover, GPI-AP...
The cellular mechanisms that ensure the selectivity and fidelity of secretory cargo protein transport from endoplasmic reticulum (ER) to Golgi are still not well understood. p24 complex acts as a specific receptor for GPI-anchored proteins by facilitating their ER exit through specialized export pathway in yeast. In parallel, can also using general exports rest with respective receptors. Here, we show biochemically associates at other receptors COPII-dependent manner, forming high-molecular...
Understanding how in eukaryotic cells thousands of proteins are sorted from each other through the secretory pathway and delivered to their correct destinations is a central issue cell biology. We have further investigated yeast two distinct types cargo into different endoplasmic reticulum (ER) exit sites (ERES) for differential ER export Golgi apparatus. used an optimized protocol that combines live dual-cargo system with 3D simultaneous multi-color high-resolution microscopy called...
Cell division produces two viable cells of a defined size. Thus, all require mechanisms to measure growth and trigger cell when sufficient has occurred. Previous data suggest model in which rate size are mechanistically linked by ceramide-dependent signals budding yeast. However, the conservation that govern control is poorly understood. In fission yeast, ceramide synthase encoded genes, Lac1 Lag1. Here, we characterize them using combination genetics, microscopy, lipid analysis. We showed...
SUMMARY Gene disruption has been dramatically facilitated by genome editing tools. Despite improvements in gene rates cultured cells, clone isolation remains routinely performed to obtain mutants, potentially leading artifacts due clonal variation cellular phenotypes. Here we report GENF, a highly efficient strategy disrupt genes without isolating clones, which can be multiplexed. Using it, obtained reliable lipidomics datasets from mutant cells being affected variances related isolation....
Glycosylphosphatidylinositol (GPI) anchoring of proteins is an essential post-translational modification in all eukaryotes that occurs at the endoplasmic reticulum (ER) and serves to deliver GPI-anchored (GPI-APs) cell surface where they play a wide variety vital physiological roles. This paper describes specialized method for purification structural analysis GPI glycan individual GPI-APs yeast. The protocol involves expression specific GPI-AP tagged with GFP, enzymatic release from cellular...
GPI anchoring is an essential post-translational modification in eukaryotes that links proteins to the plasma membrane. In this issue, Liu et al. (2023. J. Cell Biol.https://doi.org/10.1083/jcb.202208159) suggest, for first time, a regulation on demand of glycolipid precursor biosynthesis.
In eukaryotic cells, a subset of cell surface proteins is attached by the glycolipid glycosylphosphatidylinositol (GPI) to external leaflet plasma membrane where they play important roles as enzymes, receptors, or adhesion molecules. Here we present protocol for purification and mass spectrometry analysis lipid moiety individual GPI-anchored (GPI-APs) in yeast. The method involves expression specific GPI-AP tagged with GFP, solubilization, immunoprecipitation, separation electrophoresis,...
Golgi trafficking depends on the small GTPase Arf1 which, upon activation, drives assembly of different coats onto budding vesicles. Two related types guanine nucleotide exchange factors (GEFs) activate at sites. In yeast, Gea1 in cis-Golgi and Gea2 medial-Golgi to form COPI-coated vesicles for retrograde cargo sorting, whereas Sec7 generates clathrin/adaptor-coated trans-Golgi network (TGN) forward transport. A central question is how same activated protein manages assemble depending...
Lipid and protein diversity provides structural functional identity to the membrane compartments that define eukaryotic cell. This compositional heterogeneity is maintained by secretory pathway, which feeds newly synthesized proteins lipids endomembrane systems. The precise sorting of through pathway guarantees achievement their correct delivery. Although have been shown be key for mechanisms, whether how contribute this process still an open discussion. Our laboratory, in collaboration with...
Intracellular trafficking through the secretory organelles depends on transient interactions between cargo proteins and transport machinery. Cytosolic coat protein complexes capture specific luminal for incorporation into vesicles by interacting with them indirectly a transmembrane adaptor or receptor. Due to their nature, it is difficult study these ternary just using conventional native co-immunoprecipitation. To overcome this technical challenge, we have applied crosslinking assay...