Abstract:
Uridine diphosphate (UDP)-glycosyltransferases (UGTs) are important metabolizing enzymes functioning by adding a sugar moiety to a small lipophilic substrate molecule and play critical roles in drug/toxin metabolism for all realms of life. In this study, the silkworm Bombyx mori UGT33D1 gene was characterized in detail. UGT33D1 was found localized in the endoplasmic reticulum (ER) compartment just like other animal UGTs and was mainly expressed in the silkworm midgut. We first reported that UGT33D1 was important to BmNPV infection, as silencing UGT33D1 inhibited the BmNPV infection in silkworm BmN cells, while overexpressing the gene promoted viral infection. The molecular pathways regulated by UGT33D1 were analysed via transcriptome sequencing upon UGT33D1 knockdown, highlighting the important role of the gene in maintaining a balanced oxidoreductive state of the organism. In addition, proteins that physically interact with UGT33D1 were identified through immunoprecipitation and mass spectrometry analysis, which includes tubulin, elongation factor, certain ribosomal proteins, histone proteins and zinc finger proteins that had been previously reported for human UGT-interacting proteins. This study provided preliminary but important functional information on UGT33D1 and is hoped to trigger deeper investigations into silkworm UGTs and their functional mechanisms.
摘要:
尿苷二磷酸(UDP)-糖基转移酶(UGT)是重要的代谢酶,通过将糖部分添加到小的亲脂性底物分子中来发挥作用,并在所有生命领域的药物/毒素代谢中起关键作用。在这项研究中,对家蚕UGT33D1基因进行了详细的表征。与其他动物UGTs一样,UGT33D1位于内质网(ER)区室中,主要在家蚕中肠中表达。我们首次报道UGT33D1对BmNPV感染很重要,由于沉默UGT33D1抑制了家蚕BmN细胞中的BmNPV感染,而过度表达基因会促进病毒感染。通过UGT33D1敲低后的转录组测序分析了UGT33D1调节的分子途径,强调基因在维持生物体平衡的氧化还原状态中的重要作用。此外,通过免疫沉淀和质谱分析鉴定了与UGT33D1物理相互作用的蛋白质,其中包括微管蛋白,延伸率,某些核糖体蛋白,先前报道的人类UGT相互作用蛋白的组蛋白和锌指蛋白。这项研究提供了有关UGT33D1的初步但重要的功能信息,并希望引发对家蚕UGTs及其功能机制的深入研究。