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GOLGI REASSEMBLY AND STACKING PROTEIN (GRASP) PARTICIPATES IN VESICLE-MEDIATED RNA EXPORT IN CRYPTOCOCCUS NEOFORMANS
Silva, Roberta Peres da et al. | Date Issued: 2018
Author
Silva, Roberta Peres da
Martins, Sharon de Toledo
Rizzo, Juliana
Dos Reis, Flavia C. G.
Joffe, Luna S.
Vainstein, Marilene Henning
Kmetzsch, Lívia
Oliveira, Débora L.
Puccia, Rosana
Goldenberg, Samuel
Rodrigues, Marcio Lourenço
Alves, Lysangela R.
Affilliation
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Microbiologia, Imunologia e Parasitologia. São Paulo, SP, Brazil / University of Nottingham. School of Life Sciences. Nottingham, UK.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil.
Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Professor Paulo de Góes. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil.
Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia e Departamento de Biologia Molecular e Biotecnologia. Porto Alegre, RS, Brasil.
Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia e Departamento de Biologia Molecular e Biotecnologia. Porto Alegre, RS, Brasil.
Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Rio de Janeiro, RJ, Brasil
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Microbiologia, Imunologia e Parasitologia. São Paulo, SP, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil / Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Professor Paulo de Góes. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil.
Abstract
Golgi reassembly and stacking protein (GRASP) is required for polysaccharide secretion and virulence in Cryptococcus neoformans. In fungal species, extracellular vesicles (EVs) participate in the export of polysaccharides, proteins and RNA. In the present work, we investigated if EV-mediated RNA export is functionally connected with GRASP in C. neoformans using a graspΔ mutant. Since GRASP-mediated unconventional secretion involves autophagosome formation in yeast, we included the atg7Δ mutant with defective autophagic mechanisms in our analysis. All fungal strains exported EVs but deletion of GRASP or ATG7 profoundly affected vesicular dimensions. The mRNA content of the graspΔ EVs differed substantially from that of the other two strains. The transcripts associated to the endoplasmic reticulum were highly abundant transcripts in graspΔ EVs. Among non-coding RNAs (ncRNAs), tRNA fragments were the most abundant in both mutant EVs but graspΔ EVs alone concentrated 22 exclusive sequences. In general, our results showed that the EV RNA content from atg7Δ and WT were more related than the RNA content of graspΔ, suggesting that GRASP, but not the autophagy regulator Atg7, is involved in the EV export of RNA. This is a previously unknown function for a key regulator of unconventional secretion in eukaryotic cells.
Keywords
Cryptococus neoformans
RNA
Extracellular vesicles
GRASP
Atg7
Unconventional secretory pathway
 
Citation
PERES DA SILVA, R. et al. Golgi Reassembly and Stacking Protein (GRASP) Participates in Vesicle-Mediated RNA Export in Cryptococcus Neoformans. Genes (Basel), v. 9, n. 8, p. 1-17, 2018.
DOI
10.3390/genes9080400
ISSN
0378-1119