Characterization of human genes as potential chaperone with desagregase function

• Grant number: 13/10939-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): August 01, 2013
• Effective date (End): January 16, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Carlos Henrique Inacio Ramos
• Grantee: Josielle Abrahão
• Host Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 12/50161-8 - Study of the structure and function of the Hsp90 chaperone with emphasis on its role in cellular homeostasis, AP.TEM
• Associated scholarship(s): 15/13521-4 - Functional characterization of R2TP-Hsp90 complex in different organisms, BE.EP.DR

Abstract

A growing number of diseases have been correlated with the incorrect folding and protein aggregation. To protect the cells of these factors there are a group of proteins known as molecular chaperones that are intrinsically involved in protein homeostasis. These proteins help the protein folding and participate in many other cellular processes such as translocation and signaling. The most important of these chaperone functions is the capacity to disaggregate amyloid proteins. This chaperone belongs to the Hsp100 family and is called disaggregase. This chaperone is present in prokaryotes, some plants, parasites, but curiously is missing in animals. This project aims at understanding the mechanism of a human protein, now known as 'human ClpB homolog' that in recent results from our group showed function and complementation of yeast that had the disaggregase gene deleted, and to definitely establish this as a molecular chaperone disaggregase. The potential of disaggregases action in the protein aggregates can suggest their use in therapies against disease-associated conformations such as Alzheimer's and Parkinson's. The techniques to be used to achieve these results are: evaluation of expression levels in HeLa cells under conditions of stress, silencing expression 'RNAi knock-down'; heterologous expression in yeast his-tag for purification with co-interactors, and to establish a purification method of the soluble protein for structural and functional studies in vitro. Moreover, there is a possibility that the animals could have several different disaggregases with 'low activity' than a single disaggregase with 'high activity', another aim of this proposal is to test one or more candidate genes. Then, the achievement of this project has a great deal to contribute to our understanding the action mechanisms of this protein in humans, because the literature currently offers only three articles that report the function of this protein in metazoan. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ABRAHAO, JOSIELLE; AMARO, BARBARA T.; PERES, BARBARA R.; QUEL, NATALIA G.; ARAGA, ANNELIZE Z. B.; MOREA, EDNA G. O.; CANO, MARIA ISABEL N.; HOURY, WALID A.; RAMOS, CARLOS H. I.. Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity. Archives of Biochemistry and Biophysics, v. 703, MAY 30 2021. (15/13521-4, 13/10939-2, 14/25967-4, 12/50161-8, 19/11496-3)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

ABRAHÃO, Josielle. Conformational and functional characterization of AAA domain proteins through the investigation of human ANKCLP and Rvbs from Leishmania major and Saccharomyces cerevisiae. 2018. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.