Site directed mutation of residues identified as mutated on the Nek5 gene in tumored tissues and functional analysis of these mutations

• Grant number: 16/23088-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2017
• End date: January 31, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Arimi Horita
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Kinase proteins are involved in many cell processes as well as cell cycle, DNA damage response and repair, apoptosis etc. The Neks (NIMA Related Kinase) is one type of these kinase proteins involved in cell cycle regulation. In addition, many members of the Nek has their expression levels increased in cancer cells and demonstrates mutation that modifies their activities, probably contributing for the tumorigenic process. Cancer is characterized, among other facts, by accumulating several genetic changes in the DNA that can lead to gain or loss of protein function, and alteration of a normal cell processes regulation. Nek5 depletion leads to reduction of microtubules nucleation and of mitotic spindle organization and increases susceptibility to cell death. Studies performed by our group have observed an increased expression of Nek5 in tumorous cells of kidney, esophagus and thyroid. The understanding of mutations effects that occur in Nek5 gene may contribute to dealing with alternative therapies for the disease. Therefore, the central objective of this project is to perform site directed mutations on Nek5 gene that mostly occurs in cancers. Then, these mutations of Nek5 will be introduced in different types of cells and their effects will be assessed by cell cycle analysis, apoptosis, proliferation and cell localization by flow citometry and confocal microscopy. (AU)