Journal: Journal of Virology

Article Title: African swine fever virus hijacks host pyrimidine metabolism to promote viral replication

doi: 10.1128/jvi.00985-25

Figure Lengend Snippet: Endogenous aspartate synthesis regulates ASFV replication. ( A ) Exogenous aspartate deficiency does not affect the replication of ASFV (normal culture medium containing 150 µM aspartate). ( B ) Schematic diagram of endogenous synthesis pathway of aspartate. ( C ) Detection of the effect of aminooxyacetic acid hemihydrochloride (AOA) on the expression of ASFV-P30 protein using Western blotting. ( D ) RT-qPCR analysis of ASFV-B646L mRNA expression in infected cells treated with GOT1 siRNA. ( E ) Western blot and virus titration assessing the effect of GOT1 siRNA on ASFV-P30 expression and infectious virus production. ( F ) Exogenous aspartate supplementation restores ASFV-P30 protein levels suppressed by GOT1 siRNA. ( G ) RT-qPCR analysis of ASFV-B646L mRNA expression in infected cells treated with GOT2 siRNA. ( H ) Detection of the effect of GOT2 siRNA on the expression of ASFV-P30 protein expression and virus titer. For all experiments, PAMs were infected with ASFV at MOI = 1, and cell lysates were collected at 24 hpi for Western blotting. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. = no significant difference ( P ≥ 0.05).

Article Snippet: The antibodies used in this study included ASFV-P30 mouse monoclonal antibody (produced by our laboratory), β-tubulin ( M20005 ; Abmart), GOT1 (14886-1-AP; Proteintech), and GOT2 (67738-1-Ig; Proteintech).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Infection, Virus, Titration

Journal: Journal of Virology

Article Title: African swine fever virus hijacks host pyrimidine metabolism to promote viral replication

doi: 10.1128/jvi.00985-25

Figure Lengend Snippet: Temporal GOT1–GOT2 reciprocity directs aspartate flux for ASFV replication. ( A, B ) Expression levels of GOT1 and GOT2 during ASFV infection at various time points. ( C, D ) RT-qPCR and Western blot analysis of changes in GOT1 expression after GOT2 knockdown by siRNA. ( E ) Exogenous nucleoside supplementation restores ASFV-P30 protein levels suppressed by GOT1 siRNA. Unless otherwise indicated, PAMs were infected with ASFV at MOI = 1, and samples were collected at 24 hpi for Western blotting, RT-qPCR, and viral titer analysis. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. = no significant difference ( P ≥ 0.05).

Article Snippet: The antibodies used in this study included ASFV-P30 mouse monoclonal antibody (produced by our laboratory), β-tubulin ( M20005 ; Abmart), GOT1 (14886-1-AP; Proteintech), and GOT2 (67738-1-Ig; Proteintech).

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Knockdown

Journal: Journal of Virology

Article Title: African swine fever virus hijacks host pyrimidine metabolism to promote viral replication

doi: 10.1128/jvi.00985-25

Figure Lengend Snippet: Schematic diagram illustrating the mechanisms by which ASFV regulates nucleotide synthesis precursors through multiple pathways. ASFV reprograms host central carbon and nitrogen metabolism to fuel de novo pyrimidine nucleotide synthesis. First, ASFV activates the PPP, diverting glucose-derived flux to generate R5P, which is essential for nucleotide backbone synthesis. Second, ASFV upregulates the expression of the glutamine transporter SLC1A5, enhancing cellular glutamine uptake. Imported glutamine serves dual roles: (i) as a nitrogen donor for nucleotide biosynthesis, and (ii) as a carbon source, being converted to α-KG via glutaminolysis. α-KG enters the TCA cycle and is further converted by GOT1 to produce aspartate, which is required for pyrimidine ring formation. Together, these pathways ensure an adequate supply of nucleotide precursors to support ASFV DNA replication and gene expression. Viral hijacking of host metabolism thus represents a coordinated strategy to sustain robust viral replication.

Article Snippet: The antibodies used in this study included ASFV-P30 mouse monoclonal antibody (produced by our laboratory), β-tubulin ( M20005 ; Abmart), GOT1 (14886-1-AP; Proteintech), and GOT2 (67738-1-Ig; Proteintech).

Techniques: Derivative Assay, Expressing, Gene Expression

Journal: Pharmaceuticals

Article Title: Integration of Untargeted Metabolomics, Network Pharmacology, Single-Cell RNA Sequencing, and Molecular Dynamics Simulation Reveals GOT1, CYP1A2, and CA2 as Potential Targets of Huang Qin Decoction Preventing Colorectal Cancer Liver Metastasis

doi: 10.3390/ph18071052

Figure Lengend Snippet: MD trajectory analysis for CA2-HQD056, CYP1A2-HQD037, and GOT1-HQD049, including ( A ) RMSD plots, ( B ) RMSF plots, ( C ) Rg plots. ( D ) Number of hydrogen bonds. ( E ) SASA plots.

Article Snippet: Recombinant human aspartate aminotransferase (GOT1) protein (10 μg) was procured from MedChemExpress (Shanghai, China) and dissolved in 50 μL water, then diluted to a 200 μg/mL solution.

Techniques:

Journal: Pharmaceuticals

Article Title: Integration of Untargeted Metabolomics, Network Pharmacology, Single-Cell RNA Sequencing, and Molecular Dynamics Simulation Reveals GOT1, CYP1A2, and CA2 as Potential Targets of Huang Qin Decoction Preventing Colorectal Cancer Liver Metastasis

doi: 10.3390/ph18071052

Figure Lengend Snippet: Validation of the findings in MD simulation. ( A ) The 2D and 3D Gibbs free energy landscapes for CA2-HQD056, CYP1A2-HQD037, and GOT1-HQD049 are plotted against RMSD and Rg. The color scale is based on kcal/mol, where darker blue shades represent lower energy states, indicating more stable conformations. ( B ) 2D and 3D surface morphology images of the GOT1 protein and the GOT1-HQD049 complex obtained through AFM analysis.

Article Snippet: Recombinant human aspartate aminotransferase (GOT1) protein (10 μg) was procured from MedChemExpress (Shanghai, China) and dissolved in 50 μL water, then diluted to a 200 μg/mL solution.

Techniques: Biomarker Discovery

Journal: Pharmaceuticals

Article Title: Integration of Untargeted Metabolomics, Network Pharmacology, Single-Cell RNA Sequencing, and Molecular Dynamics Simulation Reveals GOT1, CYP1A2, and CA2 as Potential Targets of Huang Qin Decoction Preventing Colorectal Cancer Liver Metastasis

doi: 10.3390/ph18071052

Figure Lengend Snippet: Naringenin attenuates biological behaviors, decreases GOT1 expression, and disrupts glutamine metabolism in malignant colon cancer cells. ( A ) The CCK-8 assay was utilized to evaluate cell viability. Transwell assays quantified the number of migrated ( B ) and invaded ( C ) cells (×10). Immunofluorescence assays detected GOT1 expression ( D ) (×60), while ELISA assays measured glutamine levels in cell supernatants ( E ). *** p < 0.001.

Article Snippet: Recombinant human aspartate aminotransferase (GOT1) protein (10 μg) was procured from MedChemExpress (Shanghai, China) and dissolved in 50 μL water, then diluted to a 200 μg/mL solution.

Techniques: Expressing, CCK-8 Assay, Immunofluorescence, Enzyme-linked Immunosorbent Assay