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Identification of protein interactions involved in the regulation of carbon catabolite repression in Aspergillus nidulans

Grant number: 16/03900-0
Support type:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): June 10, 2016
Effective date (End): December 09, 2016
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal researcher:Gustavo Henrique Goldman
Grantee:Leandro Jose de Assis
Supervisor abroad: Ozgur Bayram
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Research place: National University of Ireland, Maynooth (NUI Maynooth), Ireland  
Associated to the scholarship:14/00789-6 - Functional characterization of Aspergillus nidulans phosphatases involved on glucose metabolism, BP.PD

Abstract

The fungus Aspergillus nidulans is a model organism to study the production of enzymes used in second generation biofuels production. These enzymes are used for the degradation of biomass, such as sugarcane bagasse, to produce sugars for fermentation. However, in the presence of glucose, the use of other sugars is reduced as the genes encoding enzymes involved in the usage of alternative carbon sources are subjected to carbon catabolite repression (CCR). In addition, there are other proteins involved in glucose metabolism such as phosphatases. In A. nidulans there are 7 phosphatase deletion mutants that show a reduction in the glucose assimilation. One central regulator of glycolysis is protein kinase A (PKA) that controls phosphofructokinase 1 activity thus allowing glucose metabolism to continue. The deletion of this kinase reduced glucose metabolism and released CCR, through indirectly removing the carbon catabolite repressor CreA from the nucleus and/or targeting proteins regulated by phosphatases which are involved in glucose metabolism. There is little detailed information about CreA regulation and its interaction partners. In S. cerevisiae Mig1p, the homologue of A. nidulans CreA, is controlled by phosphorylation and ubiquitination, the latter targeting it for degradation in the SCF (S-phase-kinase associated protein Cullin F-box protein) complex. The fungal Fbox proteins work in highly diverse cellular processes, participating in the removal or inactivation of specific proteins. This study proposes two aims: (i) we will try to identify proteins which physically interact with PkaA (PKA in A. nidulans) under CCR and after release the CCR. We will use a mass spectrometry-based approach after immuno-precipitation of GFP/TAP tagged proteins to discover the recruitment of proteins that interact with PkaA in these two conditions (ii) In a screening of a library of A. nidulans Fbox null mutants using xylose as a single carbon source and 2-deoxy-glucose (2DG), we have identified the mutants ”fbx23 (AN5593) and ”fbx47 (AN8909) as more sensitive and resistant to xylose+2DG, respectively. In addition, using these two identified Fbox proteins, we are going to check their possible physical interactions with targets involved in CCR. These approaches will be useful to better understanding the metabolic interplay between glucose metabolism which is subject to control by phosphatase proteins and CCR which is regulated by PkaA and Fbox proteins. (AU)

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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)
DE ASSIS, LEANDRO JOSE; SILVA, LILIAN PEREIRA; BAYRAM, OZGUR; DOWLING, PAUL; KNIEMEYER, OLAF; KRUEGER, THOMAS; BRAKHAGE, AXEL A.; CHEN, YINGYING; DONG, LIGUO; TAN, KAELING; WONG, KOON HO; RIES, LAURE N. A.; GOLDMAN, GUSTAVO H. Carbon Catabolite Repression in Filamentous Fungi Is Regulated by Phosphorylation of the Transcription Factor CreA. MBIO, v. 12, n. 1 JAN-FEB 2021. Web of Science Citations: 0.
DE ASSIS, LEANDRO JOSE; ULAS, MEVLUT; ANNICK RIES, LAURE NICOLAS; EL RAMLI, NADIA ALI MOHAMED; SARIKAYA-BAYRAM, OZLEM; BRAUS, GERHARD H.; BAYRAM, OZGUR; GOLDMAN, GUSTAVO HENRIQUE. Regulation of Aspergillus nidulans CreA-Mediated Catabolite Repression by the F-Box Proteins Fbx23 and Fbx47. MBIO, v. 9, n. 3 MAY-JUN 2018. Web of Science Citations: 4.

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