Faculty of Agriculture, University of Miyazaki.
Department of Refractory Viral Infections, National Center for Global Health and Medicine Research Institute.
Biol Pharm Bull. 2022;45(10):1559-1563. doi: 10.1248/bpb.b22-00503.
Dihydroceramide Δ4-desaturase 1 (DEGS1) enzymatic activity is inhibited with N-(4-hydroxyphenyl)-retinamide (4-HPR). We reported previously that 4-HPR suppresses severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry through a DEGS1-independent mechanism. However, it remains unclear whether DEGS1 is involved in other SARS-CoV-2 infection processes, such as virus replication and release. Here we established DEGS1 knockout (KO) in VeroE6 cells. No significant difference was observed in virus production in the culture supernatant between wild-type (WT) cells and DEGS1-KO cells, although the levels of dihydroceramide (DHCer), a DEGS1 substrate, were significantly higher in DEGS1-KO cells than WT cells. Furthermore, the virus-induced cytopathic effect was also observed in DEGS1-KO cells. Importantly, the EC value of 4-HPR in DEGS1-KO cells was almost identical to the value reported previously in WT cells. Our results indicated the lack of involvement of DEGS1 in SARS-CoV-2 infection.
二氢神经酰胺 Δ4-去饱和酶 1（DEGS1）的酶活性被 N-(4-羟基苯基）-视黄酰胺（4-HPR）抑制。我们之前报道过，4-HPR 通过一种 DEGS1 非依赖性机制抑制严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）进入。然而，DEGS1 是否参与其他 SARS-CoV-2 感染过程，如病毒复制和释放，仍不清楚。在这里，我们在 VeroE6 细胞中建立了 DEGS1 敲除（KO）。尽管 DEGS1-KO 细胞中 DEGS1 底物二氢神经酰胺（DHCer）的水平明显高于 WT 细胞，但在 WT 细胞和 DEGS1-KO 细胞的培养上清液中，病毒产量没有明显差异。此外，在 DEGS1-KO 细胞中也观察到了病毒诱导的细胞病变效应。重要的是，DEGS1-KO 细胞中 4-HPR 的 EC 值与之前在 WT 细胞中报道的值几乎相同。我们的结果表明 DEGS1 不参与 SARS-CoV-2 感染。
