Characterization of the Nek8 protein interactome using Proximity-dependent Biotin Identification (BioID)

• Grant number: 24/22038-4
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: July 01, 2025
• End date: December 31, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Kauany de Oliveira Ferreira
• Supervisor: Gagan D Gupta
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: Toronto Metropolitan University (TMU), Canada
• Associated to the scholarship: 23/15911-0 - Nek8, a serine-threonine kinase that regulates the cell cycle: functional studies of mutations found in human colon cancer, BP.MS

Abstract

The eukaryotic cell cycle is regulated by a cascade of mechanisms that ensure its integrity,in particular, the interphase (G1, S, and G2 phases) and mitosis. Protein kinases, including NIMA-related kinases (Neks), play crucial roles in cell cycle regulation. Nek8, a member of the Nek family, is involved in centrosomal organization and primary cilium formation, with mutations linked to diseases like polycystic kidney disease (PKD). Mutations in Nek8, such as T162A, G442V, and H425Y, which the mutation affects the location and function, highlighting the need for a deeper understanding of its protein interactions. This study aims to identify Nek8 interactors using BioID (proximity-dependent biotin identification), focusing on how Nek8 mutations alter its interactions and localization. The approach includes cloning the full-length Nek8 gene, creating mutations, and transfecting mammalian cells for protein analysis. Interactions will be analyzed by Liquid Chromatography-Mass Spectrometry (LC/MS), providing insights into Nek8's cellular roles. We hope this study will deepen our understanding of Nek8's function in the cell cycle and ciliogenesis, with implications for genetic disorders like nephronophthisis (NPHP) and polycystic kidney disease (PKD).