Assembly-hub function of ER-localized SNARE proteins in biogenesis of tombusvirus replication compartment

Autor: Paulina Alatriste Gonzalez, Kai Xu, Zsuzsanna Sasvari, Peter D. Nagy, Nikolay Kovalev
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Tombusvirus
Confocal Microscopy
Cell Membranes
Endoplasmic Reticulum
Virus Replication
Biochemistry
lcsh:QH301-705.5
Microscopy
biology
Chemistry
Qa-SNARE Proteins
Eukaryota
Light Microscopy
Membrane contact site
Lipids
Cell biology
Sterols
Host-Pathogen Interactions
Mitochondrial Membranes
RNA
Viral
Cellular Structures and Organelles
SNARE Proteins
Research Article
DNA Replication
lcsh:Immunologic diseases. Allergy
Saccharomyces cerevisiae Proteins
Immunology
RNA-dependent RNA polymerase
Endosomes
Saccharomyces cerevisiae
Biosynthesis
Research and Analysis Methods
Microbiology
03 medical and health sciences
Viral Proteins
Virology
Genetics
Peroxisomes
Qc-SNARE Proteins
Protein Interactions
Molecular Biology
USE1
DNA replication
Organisms
Fungi
Biology and Life Sciences
Membrane Proteins
Proteins
Cell Biology
biology.organism_classification
RNA-Dependent RNA Polymerase
Yeast
Viral Replication
030104 developmental biology
Viral replication
lcsh:Biology (General)
Parasitology
Soluble NSF attachment protein
Tomato bushy stunt virus
lcsh:RC581-607
Confocal Laser Microscopy
Zdroj: PLoS Pathogens, Vol 14, Iss 5, p e1007028 (2018)
PLoS Pathogens
ISSN: 1553-7374
1553-7366
Popis: Positive-strand RNA viruses assemble numerous membrane-bound viral replicase complexes within large replication compartments to support their replication in infected cells. Yet the detailed mechanism of how given subcellular compartments are subverted by viruses is incompletely understood. Although, Tomato bushy stunt virus (TBSV) uses peroxisomal membranes for replication, in this paper, we show evidence that the ER-resident SNARE (soluble NSF attachment protein receptor) proteins play critical roles in the formation of active replicase complexes in yeast model host and in plants. Depletion of the syntaxin 18-like Ufe1 and Use1, which are components of the ER SNARE complex in the ERAS (ER arrival site) subdomain, in yeast resulted in greatly reduced tombusvirus accumulation. Over-expression of a dominant-negative mutant of either the yeast Ufe1 or the orthologous plant Syp81 syntaxin greatly interferes with tombusvirus replication in yeast and plants, thus further supporting the role of this host protein in tombusvirus replication. Moreover, tombusvirus RNA replication was low in cell-free extracts from yeast with repressed Ufe1 or Use1 expression. We also present evidence for the mislocalization of the tombusviral p33 replication protein to the ER membrane in Ufe1p-depleted yeast cells. The viral p33 replication protein interacts with both Ufe1p and Use1p and co-opts them into the TBSV replication compartment in yeast and plant cells. The co-opted Ufe1 affects the virus-driven membrane contact site formation, sterol-enrichment at replication sites, recruitment of several pro-viral host factors and subversion of the Rab5-positive PE-rich endosomes needed for robust TBSV replication. In summary, we demonstrate a critical role for Ufe1 and Use1 SNARE proteins in TBSV replication and propose that the pro-viral functions of Ufe1 and Use1 are to serve as assembly hubs for the formation of the extensive TBSV replication compartments in cells. Altogether, these findings point clearly at the ERAS subdomain of ER as a critical site for the biogenesis of the TBSV replication compartment.
Author summary Viral replication organelles are formed in subcellular compartments during positive-strand RNA virus infections to support robust virus replication. TBSV induces multivesicular body-like structures consisting of aggregated peroxisomes. However, endoplasmic reticulum (ER) and early endosomal proteins and membranes also contribute to the biogenesis of the replication compartment. The authors show that the syntaxin 18-like Ufe1 and Use1 ER SNARE proteins, which are present in ER subdomains called ERAS (ER arrival site), are necessary for the formation of the viral replication organelles. By binding to the p33 replication protein of TBSV, Ufe1 and Use1 serve as an assembly hub for biogenesis of the replication compartment and facilitating the transfer of phospholipids and sterols to the growing sites of viral replication. The advantage of co-opting ER resident SNAREs could be that these proteins constitute very active ER subdomains (ERAS), which might be especially suitable for generation of the extensive membranous viral replication compartment.
Databáze: OpenAIRE
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