Comparative structural analyses of RPL10 tumor suppressor protein native and muted and its interaction with the c-JUN oncoprotein

• Grant number: 10/52666-4
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2011
• End date: March 31, 2014
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Regina Affonso
• Grantee: Regina Affonso
• Host Institution: Instituto de Pesquisas Energéticas e Nucleares (IPEN). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

The protein RPL10 belongs to the family of ribosomal proteins and it is highly conserved from yeast to humans. This protein is necessary for joining the 60S and 40S ribosomal subunits in the cytoplasm and to initiate of mRNA translation. Although the exact extra-ribosomal functions of RPL10 are not yet fully understood, it has been identified as a putative tumor suppressor. This protein binds JUN protein in the nucleus of the cell and it blocks the transcription of factors of cellular growth. Recent work had related the RPL10, contained mutation in the amino acids sequence, with premature ovarian failure (POF) and autism. The RPL10 gene is located at hotspot region (Xq28) containing genes involved in normal ovary development and cognitive disorders. In this case of the POF, the protein was found with five mutations in the region coder, four of which in important region of the protein structure. These alterations in the protein suggest an increase in stability of the binding with Jun protein, with a consequent increase in the occurrence of apoptosis. We already produce a human RPL10 in a soluble form in bacterial culture at temperatures of 25°C obtained 0.2 mg/mL of the pure protein. Structural analysis of the soluble protein fraction by circular dichroism showed that this protein is structured in the alpha helix and beta sheet as described in literature. The present work has as objective to understand as RPL10 protein inhibited JUN protein, and therefore to suppress the development of tumors. The strategy will be the following: (i) to express a RPL10 contend five mutations, that had been found in a patient with POF; (ii) to compare the structure this protein RPL10, native and muted; and (iii) to evaluate the capacity of binding of these two proteins with c-JUN protein. These evaluations of the RPL10s and JUN binding could be useful in the development of new diagnostic and/or therapeutic targets for tumor, POF and others diseases with which the protein intervenes. (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)

PEREIRA, LARISSA M.; SILVA, LUANA R.; ALVES, JOSEANE F.; MANN, NELIDA; SILVA, FLAVIO SOUSA; MORGANTI, LIGIA; SILVA, ISMAEL D. C. G.; AFFONSO, REGINA. A simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodies. Protein Expression and Purification, v. 101, p. 115-120, SEP 2014. (10/52666-4)