The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
| Autor: | Kevin Felipe Martho, Thiago Nunes Roberto, Joel Machado, Erika S. Nishiduka, Otávio J. B. Brustolini, Renata C. Pascon, Amanda Teixeira de Melo, Marcelo A. Vallim, Ana Tereza Ribeiro de Vasconcelos, Alexandre Keiji Tashima |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Proteomics
0301 basic medicine Transcription Genetic Science Green Fluorescent Proteins Mutant Nutritional Status Protein degradation Models Biological Article YEPD Fungal Proteins 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Gene Expression Regulation Fungal Gene chemistry.chemical_classification Multidisciplinary Methionine Virulence Calcineurin Phosphoric Monoester Hydrolases Biosynthetic Pathways Amino acid Amino Acids Sulfur Protein Transport Gene Ontology 030104 developmental biology chemistry Biochemistry Essential gene Fungal pathogenesis Cryptococcus neoformans Medicine Pathogens Sulfur 030217 neurology & neurosurgery Transcription Factors Cysteine |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-19 (2019) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-019-48433-5 |
| Popis: | Cryptococcosis is a fungal disease caused by C. neoformans. To adapt and survive in diverse ecological niches, including the animal host, this opportunistic pathogen relies on its ability to uptake nutrients, such as carbon, nitrogen, iron, phosphate, sulfur, and amino acids. Genetic circuits play a role in the response to environmental changes, modulating gene expression and adjusting the microbial metabolism to the nutrients available for the best energy usage and survival. We studied the sulfur amino acid biosynthesis and its implications on C. neoformans biology and virulence. CNAG_04798 encodes a BZip protein and was annotated as CYS3, which has been considered an essential gene. However, we demonstrated that CYS3 is not essential, in fact, its knockout led to sulfur amino acids auxotroph. Western blots and fluorescence microscopy indicated that GFP-Cys3, which is expressed from a constitutive promoter, localizes to the nucleus in rich medium (YEPD); the addition of methionine and cysteine as sole nitrogen source (SD–N + Met/Cys) led to reduced nuclear localization and protein degradation. By proteomics, we identified and confirmed physical interaction among Gpp2, Cna1, Cnb1 and GFP-Cys3. Deletion of the calcineurin and GPP2 genes in a GFP-Cys3 background demonstrated that calcineurin is required to maintain Cys3 high protein levels in YEPD and that deletion of GPP2 causes GFP-Cys3 to persist in the presence of sulfur amino acids. Global transcriptional profile of mutant and wild type by RNAseq revealed that Cys3 controls all branches of the sulfur amino acid biosynthesis, and sulfur starvation leads to induction of several amino acid biosynthetic routes. In addition, we found that Cys3 is required for virulence in Galleria mellonella animal model. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|