Identification of E3 ligases interaction partners of ubiquitin-conjugating enzyme UBE2A

• Grant number: 22/13424-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2023
• End date: December 31, 2023
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Ângela Saito
• Grantee: Estefanny Guimarães de Abreu
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Campinas , SP, Brazil

Abstract

Ubiquitination is a post-translational modification involved in many cellular processes, like mitophagy, DNA-damage repair, protein degradation by the ubiquitin-proteasome system, and synaptic transmission. UBE2A, an ubiquitin-conjugating enzyme (E2), may perform these described functions by interacting with other proteins, such as E3 ligases RAD18 and Parkin. Due to cellular functions dependent of the action of this enzyme, mutations in its gene are related to several diseases, like some cancers, neurodegenerative diseases, and X-linked intellectual disability (ID) type Nascimento. Preliminary studies demonstrated that Q93E mutation in UBE2A, found in a Brazilian family with ID type Nascimento, is associated with neuronal cytoarchitecture and synaptic transmission alterations. However, there is no knowledge about the cellular and molecular mechanisms which lead to these alterations and interaction partners involved, in special E3 ligases which act with UBE2A. Thus, the project's objective is to identify possible E3 ligases interaction partners of UBE2A in neuronal cells using TurboID and mass spectrometry. After the identification of candidates, validation and functional studies with them will be performed, like immunoprecipitation followed by Western Blot and PLA (Proximity Ligation Assay). Thereby, this project will contribute to a better understanding of molecular mechanisms associated with the UBE2A deficiency syndrome in patients with ID.