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Porcine HERC6 acts as major E3 ligase for ISGylation and is auto-ISGylated for effective ISGylation in porcine

Microb Pathog. 2024-03; 
Cuiyu Ba, Chaonan Tian, Xueying Bo, Chang Xu, Yi Zhang, Xiangyu Sun, Yuchen Nan, Chunyan Wu
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Proteins, Expression, Isolation and Analysis … Next, 25 μL of protein G agarose beads (Genscript) was added to the cell lysate and incubated for another 1 h to precipitate the immune complex. The IP supernatant was removed by … Get A Quote

摘要

Interferon-stimulated gene product 15 (ISG15) can be conjugated to substrates through ISGylation. Currently, the E3 ligase for porcine ISGylation remains unclear. Here, we identified porcine HERC5 and HERC6 (pHERC5/6) as ISGylation E3 ligases with pHERC6 acting as a major one by reconstitution of porcine ISGylation system in HEK-293?T cell via co-transfecting E1, E2 and porcine ISG15(pISG15) genes. Meanwhile, our data demonstrated that co-transfection of pISG15 and pHERC5/6 was sufficient to confer ISGylation, suggesting E1 and E2 of ISGylation are interchangeable between human and porcine. Using an immunoprecipitation based ISGylation analysis, our data revealed pHERC6 was a substrate for ISGylation and confi... More

關鍵詞

E3 ligase, HERC5, HERC6, ISGylation, Viral deubiquitinase
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