The filamentous fungus Neurospora crassa as a model organism for the functional characterization of proteins/transcription factors that regulate the carbohydrate metabolism

• Grant number: 13/24705-3
• Support type: Research Projects - Thematic Grants
• Duration: April 01, 2014 - March 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal researcher: Maria Celia Bertolini
• Grantee: Maria Celia Bertolini
• Home Institution: Instituto de Química (IQ). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Pesquisadores principais: Marcos Roberto de Mattos Fontes
• Associated grant(s): 19/05958-4 - Multi-User Equipment approved in grant 2013/24705-3: high pressure liquid chromatography system, AP.EMU; 15/25143-4 - Structural studies with proteins related to nuclear importation: visiting professor grant, AV.EXT
• Associated scholarship(s): 17/20052-6 - Functional characterization of N. crassa FLB-3 transcription factor, BP.PD; 17/10571-6 - Analysis of specificities of Importin-± from different organisms through structural and calorimetric techniques, BP.IC; 16/23536-1 - Functional characterization of protein kinases identified as regulators of glycogen metabolism in N. crassa, BP.PD; + associated scholarships 16/14384-3 - Establishment of PTP-tag method for the purification of protein complexes containing the proteins FLB-3 and RUV-1 of Neurospora crassa, BP.IC; 16/09314-6 - Structural and calorimetric studies with importin alpha of Neorospora crassa and nuclear localization sequences of transciption factores, BP.IC; 15/13814-1 - Structural characterization of transport mechanism to the cell nucleus of glycogen-related proteins metabolism in Neurospora crassa, BP.PD; 15/09935-8 - Comparison between importins-alpha of families alpha1 and alpha2 through structural and calorimetric technics, BP.IC; 15/09735-9 - Functional characterization of N. crassa FLB-3 transcription factor, BP.PD; 14/10289-0 - Structural studies with importin-± and nuclear localization sequences (NLS) of proteins related with fungi metabolism, BP.DR - associated scholarships

Abstract

Genome sequencing allows to perform regulatory studies in a deep way. In living cells genomes are not simply made of DNA and proteins, but rather of a large amount of regulatory proteins that dynamically interact with DNA and modulate gene expression. Our laboratory has been using the filamentous fungus Neurospora crassa as a model organism to study the molecular mechanisms that are involved in the glycogen metabolism regulation. Previous results showed that the metabolism of this carbohydrate is connected with signaling pathways participating in different cellular processes, which suggests the importance of the energy produced by this metabolic pathway to the biological processes. In a previous project (Proc. 08/57566-8), some transcription factors, identified as proteins regulating the glycogen metabolism in N. crassa, were functionally characterized and many of them are being studied at the present moment. In a similar way, a collection of mutant strains in protein kinases was screened and many kinases were identified as likely regulating the glycogen metabolism. This project aims to continue the characterization studies focusing in the functional and structural aspects of transcription factors and protein kinases not yet characterized in N. crassa. Considering the previous results the main objectives of this project are: 1) to investigate how some transcription factors act in the glycogen metabolism regulation, such as FLBC, a transcription factor involved in asexual development, NIT2, a transcription factor that regulates nitrogen metabolism in N. crassa and transcriptional factors annotated as hipothetical proteins. Different technical approaches will be used to achieve this goal, 2) to investigate how some transcription factor are transported to the nucleus using co-crystallization assays with the protein importin of N. crassa and Isotermal Titration Calorimetry (ITC), 3) to functionally characterize protein kinases previously identified as regulators of glycogen metabolism, 4) to characterize the connection between the circadian clock and the energy of the glycogen metabolism. In this project we expect to bring new and relevant information concerning the functional genomic of the fungus, more specifically in relation to the regulatory network existent in N. crassa. (AU)

Scientific publications

(7)

(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)

BERNARDES, NATALIA E.; FUKUDA, CINTIA A.; DA SILVA, TAINA D.; DE OLIVEIRA, HAMINE C.; DE BARROS, ANDREA C.; DREYER, THIAGO R.; BERTOLINI, MARIA CELIA; FONTES, MARCOS R. M. Comparative study of the interactions between fungal transcription factor nuclear localization sequences with mammalian and fungal importin-alpha. SCIENTIFIC REPORTS, v. 10, n. 1 JAN 29 2020. Web of Science Citations: 0.

REINA, JEFFREY; ZHOU, LIXIN; FONTES, MARCOS R. M.; PANTE, NELLY; CELLA, NATHALIE. Identification of a putative nuclear localization signal in the tumor suppressor maspin sheds light on its nuclear import regulation. FEBS OPEN BIO, v. 9, n. 7, p. 1174-1183, JUL 2019. Web of Science Citations: 1.

BAEK, MOKRYUN; VIRGILIO, STELA; LAMB, TERESA M.; IBARRA, ONEIDA; ANDRADE, JUVANA MOREIRA; GONCALVES, RODRIGO DUARTE; DOVZHENOK, ANDREY; LIM, SOOKKYUNG; BELL-PEDERSEN, DEBORAH; BERTOLINI, MARIA CELIA; HONG, CHRISTIAN I. Circadian clock regulation of the glycogen synthase (gsn) gene by WCC is critical for rhythmic glycogen metabolism in Neurospora crassa. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 116, n. 21, p. 10435-10440, MAY 21 2019. Web of Science Citations: 0.

BONI, ANA CAROLINA; AMBROSIO, DANIELA LUZ; CUPERTINO, FERNANDA BARBOSA; MONTENEGRO-MONTERO, ALEJANDRO; VIRGILIO, STELA; FREITAS, FERNANDA ZANOLLI; CORROCHER, FLAVIA ADOLFO; GONCALVES, RODRIGO DUARTE; YANG, ALLY; WEIRAUCH, MATTHEW T.; HUGHES, TIMOTHY R.; LARRONDO, LUIS F.; BERTOLINI, MARIA CELIA. Neurospora crassa developmental control mediated by the FLB-3 transcription factor. FUNGAL BIOLOGY, v. 122, n. 6, SI, p. 570-582, JUN 2018. Web of Science Citations: 4.

DE BARROS, ANDREA C.; TAKEDA, AGNES A. S.; DREYER, THIAGO R.; VELAZQUEZ-CAMPOY, ADRIAN; KOBE, BOSTJAN; FONTES, MARCOS R. M. DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway. Biochimie, v. 146, p. 87-96, MAR 2018. Web of Science Citations: 3.

BERNARDES, NATALIA E.; TAKEDA, AGNES A. S.; DREYER, THIAGO R.; CUPERTINO, FERNANDA B.; VIRGILIO, STELA; PANTE, NELLY; BERTOLINI, MARIA CELIA; FONTES, MARCOS R. M. Nuclear transport of the Neurospora crassa NIT-2 transcription factor is mediated by importin-alpha. Biochemical Journal, v. 474, n. 24, p. 4091-4104, DEC 15 2017. Web of Science Citations: 2.

VIRGILIO, STELA; CUPERTINO, FERNANDA BARBOSA; BERNARDES, NATALIA ELISA; FREITAS, FERNANDA ZANOLLI; SEKIJIMA TAKEDA, AGNES ALESSANDRA; DE MATTOS FONTES, MARCOS ROBERTO; BERTOLINI, MARIA CELIA. Molecular Components of the Neurospora crassa pH Signaling Pathway and Their Regulation by pH and the PAC-3 Transcription Factor. PLoS One, v. 11, n. 8 AUG 24 2016. Web of Science Citations: 5.