CBL - Campus del Baix Llobregat

Projecte llegit

Títol: Proving the effect of intramitochondrial enzyme filamentation on mitochondrial morphology and cell fitness

Estudiant que ha llegit aquest projecte:

Tutor/a o Cotutor/a: SEPULCRE SANCHEZ, FRANCISCO LUIS

Departament: DEAB

Títol: Proving the effect of intramitochondrial enzyme filamentation on mitochondrial morphology and cell fitness

Data inici oferta: 15-09-2025      Data finalització oferta: 15-04-2026

Estudis d'assignació del projecte:
    GR ENG SIS BIOLÒG 23

Lloc de realització:
EEABB

Paraules clau:
Autoensamblaje proteico Saccharomyces cerevisiae E. coli mutaciones compartimentación subcelular

Descripció del contingut i pla d'activitats:
We demonstrated that proteins evolve on the edge of supramolecular assembly, and that symmetric homo-oligomeric complexes are particularly prone to self-assembly into filaments.

Such reversible filamentation of homo-oligomeric complexes has also been reported for natural, wild-type complexes, particularly for metabolic enzymes during cell stress and starvation. In eukaryotes, homo-oligomeric complexes with the potential to form filaments tend to be mitochondrial proteins. However, the consequences of intramitochondrial enzyme filamentation for mitochondrial morphology and function, as well as overall cellular fitness, have not been investigated.

To address this open question, we aim to investigate the effects of filament-forming proteins targeted to mitochondria in living yeast cells. These proteins are tagged with fluorescent proteins, they are foreign to yeast cells, and they are enzymatically inactive, ensuring that any observed effects can be attributed solely to the filamentation of the complex within the yeast mitochondria. We will characterise the mitochondrial morphology of cells expressing the filament-forming complexes using fluorescence microscopy and compare it to cells expressing a version of the complex that is unable to form filaments. Additionally, we will test the fitness effects of enzyme filamentation within the mitochondria using growth curves and/or competition assays.

Taken together, by targeting filament-forming proteins to mitochondria in living yeast cells and analysing their effects on mitochondrial morphology and cellular fitness, this project will provide insights into the consequences of intramitochondrial enzyme filamentation.

Overview (resum en anglès): Protein self-assembly into complex structures such as filaments and fibrils is key to numerous cellular functions, including metabolism and stress responses. Specific point mutations can drive atypical assemblies, impairing function or causing loss of activity, which is linked to diseases like sickle-cell anemia and neurodegenerative disorders such as Alzheimer's. This study examines the effect of a filament forming protein targeted to a specific cellular compartment 1-pok fused to YFP in Saccharomyces cerevisiae cells. Four variants were used: wild type, a fiber-forming mutant (E239Y), an inactive mutant (R169M), and the double mutant. The main goal is to determine whether fibers arise within subcellular compartments and how the cell responds. To that end, plasmids with specific localization sequences were designed, PCR-amplified, cloned in E. coli, and then used to transform yeast cells. The core experimental stages plasmid design, PCR amplification, DpnI digestion, E. coli cloning, DNA purification, and Saccharomyces cerevisiae transformation-were successfully completed

Aquest projecte està relacionat amb l'adaptació al Canvi Climatic? No

Aquest projecte està relacionat amb la digitalització del seu àmbit de treball? Sí

© CBLTIC Campus del Baix Llobregat - UPC