Projects / Programmes
Understanding the role of vault ribonucleoprotein particles in the context of viral infection
| Code |
Science |
Field |
Subfield |
| 1.05.00 |
Natural sciences and mathematics |
Biochemistry and molecular biology |
|
| Code |
Science |
Field |
| 1.06 |
Natural Sciences |
Biological sciences |
Ribonucleoproteins, protein vaults, major vault protein, viral infection, HPV, HCV, immune system, antiviral response, protein compartments.
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
37987 |
PhD Fabio Lapenta |
Biochemistry and molecular biology |
Head |
2021 - 2023 |
0 |
Organisations (1)
| no. |
Code |
Research organisation |
City |
Registration number |
No. of publicationsNo. of publications |
| 1. |
1540 |
University of Nova Gorica |
Nova Gorica |
5920884000 |
0 |
Abstract
Emerging and re-emerging viral infectious diseases pose a serious threat to global healthcare. Hence, a thorough understanding of the host antiviral response and its cellular mechanisms is crucial. Vault ribonucleoprotein organelles are large and highly conserved macromolecular complexes, which have been recently associated with the immunological response to infection of several human viruses. Vaults are nano-compartments connected to multiple cellular pathways: nucleocytoplasmic transport, multidrug resistance, apoptosis and innate immunity. To date, however, their exact role in the cellular milieu remains unclear. With the proposed project we intend to study the function of vault particles in the context of viral infection. In accordance with the current state-of-the-art, we selected two human oncogenic viruses, hepatitis B virus (HBV) and human papilloma virus (HPV) as model viral agents. We propose a bipartite research project that will cover i) the investigation of vault’s cellular role, regulation and network of interaction with other cellular components in the context of viral infection; ii) in addition to a comprehensive structural and biophysical investigation of the ribonucleoprotein particle. Vault’s expression, localization and overall cellular role will be examined before, during and in response to HBV and HPV infection of human cell lines, employing safe replication-incompetent pseudo viral particles. Expression of reporter-based systems and immunofluorescence microscopy will provide information regarding the biology of vaults and their localization. The time-course of vault’s gene expression and protein translation in response to viral infection will be tracked via flow cytometry and immunochemistry; cell lines depleted of vault components will be evaluated in parallel. Following, proteomic analysis of the vault’s interactome will be used to study the interaction network of vault particles in physiological conditions compared to cells undergoing viral infection. Simultaneously, we will study the mechanism of action, binding sites and structural properties of this organelle expressing its major protein component in vitro. Site-specific mutagenesis will be employed to identify the amino acids involved in vault’s molecular interactions. Binding analysis will be performed on proteins expressed in yeast and bacteria, followed by structural studies performed via small-angle X-ray scattering and electron microscopy on purified protein preparations. Ultimately, we believe the proposed research project will bring about a better understanding of this ribonucleoprotein complex, especially in connection to viral infection and the host immune system, providing new answers and directions to the scientific community involved in the fields of virology, antiviral applications and protein compartments.
