Rab1b-GBF1-ARFs mediated intracellular trafficking is required for classical swine fever virus replication in swine umbilical vein endothelial cells.

Abstract

Classical swine fever virus (CSFV), a plus-sense RNA virus, utilizes host intracellular membrane organelles for its replication. Our previous studies have shown that disruption of the intracellular membrane-trafficking events can inhibit CSFV replication. However, the underlying mechanism of this process in CSFV infection has not been elucidated. To determine the role of Golgi-associated anterograde and retrograde trafficking in CSFV replication, we revealed the effect of vesicular transport between Golgi and ER inhibitors Brefeldin A (BFA) and 2,2-methyl-N-(2,4,6,-trimethoxyphenyl) dodecanamide (CI-976), the GBF1 inhibitor golgicide A (GCA) on virus production. Our results showed that disruption of vesicular trafficking by BFA, CI-976, and GCA significantly inhibited CSFV infection. Subsequent experiments revealed that knockdown of Rab1b by lentiviruses and negative-mutant Rab1b-N121I transfection inhibited CSFV infection. Furthermore, we showed that the Rab1b downstream vesicular component effectors GBF1, and class I and class II ADP-ribosylation factors (ARFs) were also involved in virus replication. In addition, confocal microscopy assay showed that CSFV infection disrupted the Golgi apparatus resulting in extended Golgi distribution around the nucleus. We also showed that cell secretory pathway, measured using Gaussia luciferase flash assay, was blocked in CSFV infected cells. Taken together, these findings demonstrate that CSFV utilizes Rab1b-GBF1-ARFs mediated trafficking to promote its own replication. These findings also provide new insights into the intracellular trafficking pathways utilized for CSFV life cycle.

• Authors: Liang Zhang, Tao Wang, Mengzhao Song, Mingxing Jin, Shanchuan Liu, Kangkang Guo, Yanming Zhang
• Journal: Veterinary microbiology (Vet Microbiol) Vol. 246 Pg. 108743 (Jul 2020) ISSN: 1873-2542 [Electronic] Netherlands
• PMID: 32605744 (Publication Type: Journal Article)
• Copyright: Copyright © 2020 Elsevier B.V. All rights reserved.

Chemical References

• Guanine Nucleotide Exchange Factors
• Protein Synthesis Inhibitors
• Brefeldin A
• rab1 GTP-Binding Proteins

Topics

• Animals
• Brefeldin A (pharmacology)
• Classical Swine Fever Virus (genetics, physiology)
• Endothelial Cells (virology)
• Guanine Nucleotide Exchange Factors (genetics, metabolism)
• Protein Synthesis Inhibitors (pharmacology)
• Protein Transport
• Swine
• Umbilical Veins (cytology)
• Virus Replication (drug effects)
• rab1 GTP-Binding Proteins (genetics, metabolism)