Identification and functional studies of NEK1 and NEK2 interaction partners and substrates on the human centrosome and their relevance in ciliopathies

• Grant number: 20/16265-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2021
• End date: July 31, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Ivan Rosa e Silva
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 17/03489-1 - From functional studies to searching for new inhibitors for cancer: exploring kinases that regulate the cell cycle of the human NEK family, AP.TEM

Abstract

Centrioles assemble the centrosome, the microtubule organizing centre essential for cell division in proliferating cells. Centrioles also template cilia, cell organelles that act as antenna in quiescent cells and can also generate fluid flow. Because of their essential functions in human life, centrioles are highly regulated throughout the cell cycle. Among the least studied centriole cycle regulators are the members of the never-in-mitosis A-related kinases (NEKs) family. At least 8 NEK isoforms have been related to the centriole. In particular, NEK2 is a centrosome resident throughout the cell cycle. This project aims to identify the phosphorylation substrates of NEK1 and NEK2 in the human centrosome using mass spectrometry. We will analyse amino acid sequence conservation and use co-IP and sub-cellular localization experiments in order to validate our findings. Phosphomimetic and phosphodeficient mutants of selected interaction partners will be tested in vivo for their function in cell cycle progression and ciliogenesis. We further aim to characterize the molecular details of NEK2 interaction with centriolar sub-distal appendage components such as ODF2 in a likely essential regulation step in the G2/M transition. Therefore, our data will provide new insights into the regulation of the centrosome cycle by NEKs in human cells determining molecular details of this process. (AU)