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Structural studies with the importin-alpha from Neurospora crassa and nuclear localization sequences (NLS) peptides

Grant number: 12/16371-5
Support type:Scholarships in Brazil - Master
Effective date (Start): October 01, 2012
Effective date (End): July 31, 2014
Field of knowledge:Biological Sciences - Biophysics - Molecular Biophysics
Principal researcher:Marcos Roberto de Mattos Fontes
Grantee:Natália Elisa Bernardes
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated research grant:08/57566-8 - The Neurospora crassa filamentous fungus as a model organism for the identification and characterization of new proteins/transcription factors regulating the glycogen metabolism, AP.TEM

Abstract

The communication between the cytoplasm and nucleus occurs through transport mechanisms that allow the passage of molecules through the nuclear envelope (Lange et al., 2007; Stewart, 2007). Among the routes of transport Known that allow the transport of macromolecules into or out of the nucleus through the recognition of specific signaling sequences, the so-called Classical Nuclear Import Pathway is the most well characterized. In this way, the importinz-alpha acts in the identification of proteins to be transported to the nucleus by the recognition of nuclear localization sequences (NLS). The filamentous fungus Neurospora crassa is widely used in biochemical and molecular biology since the twentieth century (Galagan, 2003). The study of proteins of the fungus Neurospora crassa aims to allocate the biological functions of hypothetical proteins related to this metabolism. In this context, the importin-alpha appears as a model to test the function of these hypothetical proteins and this project aims to obtain the protein structure through crystallographic studies. Considering the previous studies with this protein in which was not possible suitable crystals for X-ray diffraction, will be also conducted protein crystallization experiments with NLS peptides, including the SV40 NLS peptide and protein NCU03482. As shown recently by us (Takeda et al 2012), the formation of complexes with importin-± confers stability to the protein. The studies proposed in this project will be conducted in collaboration with Prof. Maria Célia Bertolini from the Institute of Chemistry, UNESP - Araraquara campus. The project is directly related to the projects (SMOLBnet 2) - proc. 2010/51889-0 and the thematic project proc.08/57566-8. (AU)

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