Our main focus is to understand the role played by RNA molecules in protein networks. Characterizing protein-RNA associations is key to unravel the complexity and functionality of mammalian genomes and will open up therapeutic avenues for the treatment of a broad range of human disorders. We aim to discover the involvement of RNA molecules in regulatory networks controlling protein production and we are interested in understanding mechanisms whose alteration lead to aberrant aggregation, such as the one observed in amyloid diseases. We have recently observed that interaction between proteins and RNAs induce feedback loops that are crucial in protein homeostasis. We also found that specific RNAs promote phase separation of proteins in the cytosol and nucleus and we are designing synthetic RNA molecules to manipulate the formation of complexes.
In our group, the research is highly interdisciplinary, involves a number of international collaborators and focuses on structural biology and the uncharted territory of non-coding RNAs involved in transcriptional and translational regulation (e.g., X-chromosome inactivation) as well as human disorders (e.g., Parkinson’s disease). We aim to identify regulatory mechanisms controlling protein production and contributing to cell homeostasis.
Examples of our research activity include the following lines:
On the Mechanism of Genetic Inheritance Associated with the X Chromosome. We analyzed the process of inactivation of the X chromosome and in particular, the role of the RNA molecule called Xist (X-Inactive-Specific-Transcript), its main regulator. We studied the mechanism of action, structure and interactions of the Xist molecule. We observed that Xist acts as a "scaffold", it provides scaffolding and at the same time attracts lots of proteins to organize the "silencing" of the X chromosome. The interaction network is so great that Xist and its partner proteins form a structure that resembles a corpuscle, conceptually similar to a drop of oil in water.
RNA structure drives proteins crazy. We found that messenger RNA could act as a solubilizer, blocking the formation of protein aggregates that are potentially toxic to our organisms. In particular, we observed that the transcript coding for Heat Shock Protein 70 (HSP70) interacts with many proteins and has a strong effect on protein aggregation. We experimentally demonstrated for the first time that, under conditions of stress, HSP70 mRNA has the ability to promote the removal of the protein aggregates that are responsible for serious neuro-degenerative diseases such as Alzheimer’s and Amyotrophic Lateral Sclerosis.
We use computers but we also have in the lab:
- Fast protein liquid chromatography system for protein purification
- Multi-functional spectrophotometer for structural and kinetics studies
- Spectropolarimeter for circular and linear dichroism analysis
- Multi-well plate fluorescence reader with a temperature controller for aggregation studies
- Bio-layer interferometer for the study of interaction kinetics between proteins and oligonucleotides
- Annalisa Pastore
- Andrea Cerase
- Antonios Deligiannakis
- George Paliouras
- Michele Vendruscolo
- Mitch Guttman