Structural characterization of transport mechanism to the cell nucleus of glycogen-related proteins metabolism in Neurospora crassa

• Grant number: 15/13814-1
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): September 01, 2015
• Effective date (End): August 31, 2019
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal researcher: Marcos Roberto de Mattos Fontes
• Grantee: Andréa Coelho de Barros
• Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated research grant: 13/24705-3 - The filamentous fungus Neurospora crassa as a model organism for the functional characterization of proteins/transcription factors that regulate the carbohydrate metabolism, AP.TEM

Abstract

There are several nuclear transport pathways that transport of macromolecules into or out of the nucleus through recognition of specific signal sequences, the classic nuclear protein import pathway is the best characterized of these transport cycles. In this process, the structural study of Importin-± (IMP±) is the key for your understanding. This protein was recently crystallized and its structure was elucidated by our research group. Biophysics experiments with NcImp± protein have also been performed by us and showed that the protein retains some features present in Mus Musculus and Saccharomyces cerevisiae isoforms that already had their crystallographic structure previously solved. However, the NcIMP± displays amino acids not conserved near its IBB domain, which may be related to specificity of this protein. Thus, we are proposing in this project: a) co-crystallize and elucidate the structure of NcIMP± with peptide (NLS) of the protein involved in the regulation of glycogen metabolism; b) to perform affinity experiments to quantify the interactions between NcIMP± and NLS peptides using calorimetric techniques; c) to express, purify and preliminarily characterize proteins involved in glycogen metabolism from N. crassa.