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Genes 2018, 9(8), 400;

Golgi Reassembly and Stacking Protein (GRASP) Participates in Vesicle-Mediated RNA Export in Cryptococcus neoformans

Departamento de Microbiologia, Imunologia e Parasitologia da Escola Paulista de Medicina-UNIFESP, São Paulo, SP 04023-062, Brazil
School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK
Instituto Carlos Chagas, Fundação Oswaldo Cruz, Fiocruz-PR, Curitiba, PR 81310-020, Brazil
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-901, Brazil
Centro de Desenvolvimento Tecnológico em Saúde (CDTS), Fundação Oswaldo Cruz, Rio de Janeiro, RJ 21040-900, Brazil
Centro de Biotecnologia e Departamento de Biologia Molecular e Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91501-970, Brazil
These authors contributed equally to this work.
Author to whom correspondence should be addressed.
Received: 1 July 2018 / Revised: 25 July 2018 / Accepted: 31 July 2018 / Published: 8 August 2018
(This article belongs to the Special Issue Fungal Pathogenesis in Humans: The Growing Threat)
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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. View Full-Text
Keywords: Cryptococus neoformans; RNA; extracellular vesicles; GRASP; Atg7; unconventional secretory pathway Cryptococus neoformans; RNA; extracellular vesicles; GRASP; Atg7; unconventional secretory pathway

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Peres da Silva, R.; Martins, S.T.; Rizzo, J.; dos Reis, F.C.G.; Joffe, L.S.; Vainstein, M.; Kmetzsch, L.; Oliveira, D.L.; Puccia, R.; Goldenberg, S.; Rodrigues, M.L.; Alves, L.R. Golgi Reassembly and Stacking Protein (GRASP) Participates in Vesicle-Mediated RNA Export in Cryptococcus neoformans. Genes 2018, 9, 400.

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