| Full text | |
| Author(s): |
de Melo, Amanda Teixeira
[1]
;
Martho, Kevin Felipe
[1]
;
Roberto, Thiago Nunes
[1]
;
Nishiduka, Erika S.
[2]
;
Machado Jr, Joel
;
Brustolini, Otavio J. B.
[3]
;
Tashima, Alexandre K.
[2]
;
TerezaVasconcelos, Ana
[3]
;
Vallim, Marcelo A.
[4]
;
Pascon, Renata C.
[4]
Total Authors: 10
|
| Affiliation: | [1] Univ Fed Sao Paulo, Campus Diadema, Sao Paulo, SP - Brazil
[2] Univ Fed Sao Paulo, Escola Paulista Med, Dept Bioquim, Campus Sao Paulo, Sao Paulo, SP - Brazil
[3] Labinfo Lab Bioinformat, LNCC, Petropolis, RJ - Brazil
[4] Machado Jr, Jr., Joel, Univ Fed Sao Paulo, Campus Diadema, Sao Paulo, SP - Brazil
Total Affiliations: 4
|
| Document type: | Journal article |
| Source: | SCIENTIFIC REPORTS; v. 9, AUG 15 2019. |
| Web of Science Citations: | 2 |
| Abstract | |
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. (AU) | |
| FAPESP's process: | 16/14542-8 - Genetic characterization of the regulatory elements of the biosynthesis and uptake of amino acids and their role in virulence of Cryptococcus neoformans. |
| Grantee: | Renata Castiglioni Pascon |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 18/20390-1 - Study of the regulatory elements control the amino acids uptake in Cryptococcus neoformans |
| Grantee: | Kevin Felipe Cruz Martho |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 17/20106-9 - Peptidomics of Brazilian snake and spider venoms |
| Grantee: | Alexandre Keiji Tashima |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 15/04400-9 - The role of autophagy in Cryptococcus neorformas high temperature (37°C) growth and virulence |
| Grantee: | Marcelo Afonso Vallim |
| Support Opportunities: | Regular Research Grants |