Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production.

Autor:	Cardoso RS; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Tavares LA; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Jesus BLS; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Criado MF; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., de Carvalho AN; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Souza JP; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Bedi S; Department of Microbiology and Immunology, University Michigan Medical School, Ann Arbor, Michigan, USA., de Souza MM; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Silva ML; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Lanfredi GP; Department of Biochemistry, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Vitti BC; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Scudero OB; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Faça VM; Department of Biochemistry, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Ono A; Department of Microbiology and Immunology, University Michigan Medical School, Ann Arbor, Michigan, USA., Ventura AM; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., daSilva LLP; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil., Arruda E; Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, Brazil eaneto@fmrp.usp.br.
Jazyk:	angličtina
Zdroj:	MBio [mBio] 2020 Sep 29; Vol. 11 (5). Date of Electronic Publication: 2020 Sep 29.
DOI:	10.1128/mBio.01869-20
Abstrakt:	Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites. IMPORTANCE The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process. (Copyright © 2020 Cardoso et al.)
Databáze:	MEDLINE
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