hNek10 protein kinase functional and interaction profile in DNA damage repair

• Grant number: 13/11160-9
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: September 01, 2013
• End date: February 28, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Priscila Ferreira Papa
• Supervisor: Joan Roig Amorós
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Institution abroad: Institute for Research in Biomedicine (IRB), Spain
• Associated to the scholarship: 10/15262-2 - Cloning and expression of human protein kinase Nek10 structural and functional studies aimed at molecular and cellular, BP.DD

Abstract

Genomic instability and mutations are considered one of the Hallmarks of cancer. The diversity of DNA damage repair pathways can preventing that the errors could be passed to daughter cells maintaining the cell normal state. In this sense, the Nek family members are involved in these pathways through their signs and catalytic activities. One of these members is hNek10 already characterized by its mediation in response to UV stimuli. Through preliminary results of two-hybrid was possible to identify a prey member of the cohesin complex, already described as a participant in DNA repair. Thus, it became important to investigate the functional profile of hNek10 in repair pathways in collaboration with the group of Dr. Joan Roig. In addition, we will carry out the treatment of cells with DNA damaging agents at the Travis Stracker[s research group. The results obtained in analysis of mass spectrometry, microscopy and RT-PCR data will provide new interactome, location and expression of hNek10. This result will be compared with cells presenting hNek10 silenced, mutated or overexpressed. Thus, western blot analysis, confocal microscopy and time-lapse analysis will allow to analyze the hNek10 protein with known proteins in the repair pathways and interactors. In addition, flow cytometric analyzes will separate populations of cell cycle diferences, if it occurs any mitotic stops at the checkpoints. From these new data it will be possible to characterize and investigate the functionally hNek10 in this particular cellular process. (AU)