UBXN1 interacts with the S1 protein of transmissible gastroenteritis coronavirus and plays a role in viral replication

Abstract

Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs and is associated with high morbidity and mortality in sucking piglets. S1 is one of two protein domains in the spike (S) glycoprotein and is responsible for enteric tropism, sialic acid recognition, and host receptor binding. Although there has been extensive research on the S1 protein of TGEV, little is known about the intracellular role of TGEV-S1. In the present study, we used yeast two-hybrid screening of a cDNA library from porcine intestinal cells to identify proteins that interact with TGEV-S1. Among 120 positive clones from the library, 12 intracellular proteins were identified after sequencing and a BLAST search. These intracellular proteins are involved in protein synthesis and degradation, biological signal transduction, and negative control of signaling pathways. Using a glutathione- S -transferase (GST) pulldown assay and Co-IP, we found that UBXN1 interacts with the S1 protein. Here, we observed that TGEV infection led to increased UBXN1 expression levels during the late phase of infection in IPEC-J2 cells. Inhibition of UBXN1 in IPEC-J2 cells via siRNA interference significantly decreased the viral titer and downregulated the expression of S1. UBXN1 overexpression significantly increased the viral copy number. Additionally, we provided data suggesting that UBXN1 negatively regulates IFN-β expression after TGEV infection. Finally, our research indicated that UBXN1 plays a vital role in the process of TGEV infection, making it a candidate target for the development of a novel antiviral method.

HSP90AB1 Is a Host Factor Required for Transmissible Gastroenteritis Virus Infection

“…TGEV is an important enteropathogen that causes diarrhea, vomiting, and dehydration in pigs of various ages. To date, host factors, including porcine aminopeptidase N (APN) [32], TRF1 [12], TMEM41B [13], EIF4A2 [14,15], and UBXN1 [14,15], play a positive role in TGEV infection. Moreover, some host protein inhibitors, including the receptor tyrosine kinase inhibitor A9 [16] and the Na+/K+-ATPase inhibitor ouabain [17], can inhibit TGEV infection to various extents.…”

“…Transmembrane protein 41B (TMEM41B) is a host factor required for TGEV replication and contributes to the formation of coronavirus replication organelles [13]. Eukaryotic translation initiation factor 4-alpha (EIF4A2) and UBX domaincontaining protein 1 (UBXN1) play roles in viral replication and can interact with the membrane and spike protein of TGEV, respectively [14,15]. Moreover, increasing studies have indicated that small molecule inhibitors can inhibit TGEV replication by regulating intracellular signaling pathways.…”

Proteomic and phosphorylated proteomic landscape of injured lung in juvenile septic rats with therapeutic application of umbilical cord mesenchymal stem cells

“…Arrdc1 is involved in a variety of processes including cellular protein metabolism, extracellular vesicle biogenesis, and negative regulation of the Notch signaling pathway (30).The down-regulated differential protein AIF1 has been identified as an up-regulated protein of infection (31). The ubiquitin-regulated X (UbX) protein UbXN1, which contains a ubiquitin-associated domain, is a negative regulator of nuclear factor-kB (NF-kB) signaling (32,33). It is suggested that the immune inflammatory response is important in the proteome of septic lung tissue.…”

The Roles of Apoptosis in Swine Response to Viral Infection and Pathogenesis of Swine Enteropathogenic Coronaviruses

“…E protein promotes TGEV maturation in the secretory pathway (65). S1 and M proteins play a role in viral replication (66,67). In addition, M protein can affect TGEV-induced IFN-α production (68).…”