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[Objective] To investigate the specific role and molecular mechanism of the host protein protein disulfide isomerase A4 (PDIA4) during bovine viral diarrhea virus (BVDV) infection, thus providing a theoretical basis for elucidating the role of this protein in viral replication and developing novel prevention and control strategies. [Methods] Western blotting and qPCR were employed to analyze PDIA4 expression in Madin-Darby bovine kidney (MDBK) cells following BVDV infection. The expression levels of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) and sequestosome 1 (p62) were measured by Western blotting and laser confocal microscopy. MDBK cell lines with pdia4 knockdown and overexpression were constructed. The effects of pdia4 on LC3 and p62 expression were examined, and the autophagic flux was evaluated by a tandem GFP-mRFP-LC3 reporter system. Cells were subjected to starvation or treated with bafilomycin A1 (BafA1), followed by BVDV infection. Viral replication was assessed by measuring BVDV mRNA levels through qPCR and double-stranded RNA (dsRNA) levels through immunofluorescence staining. [Results] BVDV infection significantly upregulated the endogenous expression of PDIA4. The LC3 level increased progressively with infection time, whereas the p62 level showed an initial increase followed by a decrease. The cell lines with pdia4 knockdown and overexpression were successfully established. pdia4 knockdown significantly increased both LC3 and p62 levels, whereas pdia4 overexpression increased the LC3 level and decreased the p62 level. Under starvation or BafA1 treatment, pdia4 knockdown inhibited BVDV replication, whereas pdia4 overexpression promoted BVDV replication. [Conclusion] BVDV infection upregulates host PDIA4 expression. PDIA4 promotes BVDV replication by activating the autophagic flux through regulation of LC3 and p62. These findings provide new insights into the pathogenic mechanism of BVDV and offer a theoretical basis for developing targeted antiviral strategies.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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