| Texto completo | |
| Autor(es): Mostrar menos - |
de Assis, Leandro Jose
[1]
;
Silva, Lilian Pereira
[2, 3]
;
Bayram, Ozgur
[4]
;
Dowling, Paul
[4]
;
Kniemeyer, Olaf
[5]
;
Krueger, Thomas
[5]
;
Brakhage, Axel A.
[5]
;
Chen, Yingying
[6]
;
Dong, Liguo
[6]
;
Tan, Kaeling
[6]
;
Wong, Koon Ho
[6]
;
Ries, Laure N. A.
[1]
;
Goldman, Gustavo H.
[2, 3]
Número total de Autores: 13
|
| Afiliação do(s) autor(es): | [1] Univ Exeter, MRC Ctr Med Mycol, Exeter, Devon - England
[2] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto - Brazil
[3] Tech Univ Munich, Inst Adv Study, Garching - Germany
[4] Maynooth Univ, Biol Dept, Maynooth, Kildare - Ireland
[5] Friedrich Schiller Univ, Inst Microbiol, Dept Mol & Appl Microbiol, Leibniz Inst Nat Prod Res & Infect Biol, Jena - Germany
[6] Univ Macau, Fac Hlth Sci, Macau - Peoples R China
Número total de Afiliações: 6
|
| Tipo de documento: | Artigo Científico |
| Fonte: | MBIO; v. 12, n. 1 JAN-FEB 2021. |
| Citações Web of Science: | 0 |
| Resumo | |
Filamentous fungi of the genus Aspergillus are of particular interest for biotechnological applications due to their natural capacity to secrete carbohydrate active enzymes (CAZy) that target plant biomass. The presence of easily metabolizable sugars such as glucose, whose concentrations increase during plant biomass hydrolysis, results in the repression of CAZy-encoding genes in a process known as carbon catabolite repression (CCR), which is undesired for the purpose of largescale enzyme production. To date, the C2H2 transcription factor CreA has been described as the major CC repressor in Aspergillus spp., although little is known about the role of posttranslational modifications in this process. In this work, phosphorylation sites were identified by mass spectrometry on Aspergillus nidulans CreA, and subsequently, the previously identified but uncharacterized site S262, the characterized site S319, and the newly identified sites S268 and T308 were chosen to be mutated to nonphosphorylatable residues before their effect on CCR was investigated. Sites S262, S268, and T308 are important for CreA protein accumulation and cellular localization, DNA binding, and repression of enzyme activities. In agreement with a previous study, site S319 was not important for several here-tested phenotypes but is key for CreA degradation and induction of enzyme activities. All sites were shown to be important for glycogen and trehalose metabolism. This study highlights the importance of CreA phosphorylation sites for the regulation of CCR. These sites are interesting targets for biotechnological strain engineering without the need to delete essential genes, which could result in undesired side effects. IMPORTANCE In filamentous fungi, the transcription factor CreA controls carbohydrate metabolism through the regulation of genes encoding enzymes required for the use of alternative carbon sources. In this work, phosphorylation sites were identified on Aspergillus nidulans CreA, and subsequently, the two newly identified sites S268 and T308, the previously identified but uncharacterized site S262, and the previously characterized site S319 were chosen to be mutated to nonphosphorylatable residues before their effect on CCR was characterized. Sites S262, S268, and T308 are important for CreA protein accumulation and cellular localization, DNA binding, and repression of enzyme activities. In agreement with a previous study, site S319 is not important for several here-tested phenotypes but is key for CreA degradation and induction of enzyme activities. This work characterized novel CreA phosphorylation sites under carbon catabolite-repressing conditions and showed that they are crucial for CreA protein turnover, control of carbohydrate utilization, and biotechnologically relevant enzyme production. (AU) | |
| Processo FAPESP: | 16/03900-0 - Identificação de interações proteicas na regulação da repressão catabólica do carbono em Aspergillus nidulans |
| Beneficiário: | Leandro Jose de Assis |
| Modalidade de apoio: | Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado |
| Processo FAPESP: | 16/07870-9 - A influência de proteínas quinases ativadas por mitógenos (MAPK) na expressão de determinantes genéticos importantes para a virulência de Aspergillus fumigatus |
| Beneficiário: | Gustavo Henrique Goldman |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 14/00789-6 - Caracterização funcional de fosfatases de Aspergillus nidulans envolvidas no metabolismo da glicose |
| Beneficiário: | Leandro Jose de Assis |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 17/14159-2 - O papel do metabolismo da lactose e do acetato na virulência de Aspergillus fumigatus |
| Beneficiário: | Laure Nicolas Annick Ries |
| Modalidade de apoio: | Auxílio à Pesquisa - Jovens Pesquisadores |