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Study of the selenocysteine incorporation pathway: understanding the macromolecular interaction mechanism

Grant number: 16/20977-7
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): May 01, 2017
Effective date (End): February 28, 2019
Field of knowledge:Biological Sciences - Biophysics
Principal Investigator:Otavio Henrique Thiemann
Grantee:Jessica Fernandes Scortecci
Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Naegleria fowleri, commonly known as "brain-eating amoeba", is the only species of Naegleria known to infect humans, resulting in the condition known as Primary Amebic Meningoencephalitis (PAM). Despite its medical relevance it is still an organism with little attention. Our group described the presence of the selenocystein synthesis pathway in N. gruberi, a nonpathogenic species used as a model system for Naegleria studies and has contributed to elcidade this pathway both in eukaryotes as well as in prokaryotes. Selenocysteine (Sec) is incorporated into nascent protein in a UGA codon by a co-translational event both in prokaryotes and in eukaryotes. In bacteria, the Sec synthesis pathway is composed of the enzymes Selenocysteine Synthase (SELA), a Specific Elongation Factor (SELB), Selenophosphate Synthetase (SELD), seryl-tRNA synthetase (SerRS) and a Selenocysteine lyase (CSDB). The Sec synthesis and incorporation route also depends on two RNAs; a specific tRNA (tRNASec) and a specific sequence in the mRNA (Selenocisteínas Insertion Sequence - SECIS), signaling to correct incorporation of Sec codon UGA. In eukaryotes, this pathway differs by the presence of O-fosfoseril-tRNASec Kinase (PSTK) and Selenocisteinil-tRNASec Synthase (SepSecS), replacing SelA, and the presence of SECIS binding proteins (SBP2) plus a factor Specific elongation factor (EFSec). This proposal aims at biochemical and structural studies of the proteins SELD CSDB of Escherichia coli and SBP2 of N. gruberi, to establish the molecular interactions that determine the specificity of these enzymes. Testing of interaction between CSDB-SelD and SBP2-SECIS will be conducted to determine the interaction constants through spectroscopic and calorimetric techniques. SBP2 in vivo immunofluorescence tests will be performed. Moreover, crystallization tests of different complexes and X-ray small angle scattering (SACS) will be performed to obtain structural models. Thus, this project aims to increase our knowledge and understanding of the molecular interactions present in selenocysteine synthesis route, being of relevance in the global understanding of the molecular interaction determinants between protein-protein and protein-RNA. (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)
BALASCO SERRAO, VITOR HUGO; SILVA, IVAN ROSA; ALVES DA SILVA, MARCO TULIO; SCORTECCI, JSSICA FERNANDES; FERNANDES, ADRIANO DE FREITAS; THIEMANN, OTAVIO HENRIQUE. The unique tRNA(Sec) and its role in selenocysteine biosynthesis. Amino Acids, v. 50, n. 9, p. 1145-1167, SEP 2018. Web of Science Citations: 0.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.
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