Proteomic analysis of Atg8-dependent recruitment of phagosomal proteins in the enteric protozoan parasite Entamoeba histolytica.
Proteomic analysis of Atg8-dependent recruitment of phagosomal proteins in the enteric protozoan parasite Entamoeba histolytica.
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DOI：
10.3389/fcimb.2022.961645
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发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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自噬是真核生物中一种保守的降解系统。在肠道原生动物寄生虫溶组织内阿米巴（Entamoeba histolytica）（人类阿米巴病的病原体）中，Atg 8不仅参与自噬本身，还参与其他膜交通相关途径，如吞噬体生物发生。我们以前报道过反义小RNA介导的转录基因沉默（GS）抑制atg 8基因表达导致生长迟缓，内吞作用延迟，内体和吞噬体酸化减少。在这项研究中，为了更好地了解Atg 8在吞噬和胞吞作用中的作用，我们对从野生型和atg 8-gs菌株中分离的吞噬体进行了比较蛋白质组学分析。我们发现，与对照菌株相比，从atg 8-gs菌株分离的吞噬体中分别检测到127和107种蛋白质，其丰度分别为对照菌株的1/1.5或1/1.5。在127种蛋白质中，其丰度从atg 8-gs在吞噬体中减少，鉴定了一组与脂肪酸代谢、吞噬作用和内质网（ER）稳态相关的蛋白质。各种溶酶体水解酶和它们的受体也倾向于被atg 8-gs从吞噬体中排除，加强了Atg 8参与吞噬体酸化和消化的概念。相反，在atg 8-gs菌株吞噬体中丰度增加的107种蛋白中，核糖体相关蛋白和代谢物互变酶富集。我们进一步研究了几种代表性蛋白质的定位，包括腺苷酸环化酶相关蛋白和质膜钙泵，这两种蛋白质都被证明是通过Atg 8依赖性机制被招募到吞噬体和trogosome的。综上所述，我们的研究为进一步阐明E.溶组织剂
Autophagy is one of the bulk degradation systems and is conserved throughout eukaryotes. In the enteric protozoan parasite Entamoeba histolytica, the causative agent of human amebiasis, Atg8 is not exclusively involved in autophagy per se but also in other membrane traffic-related pathways such as phagosome biogenesis. We previously reported that repression of atg8 gene expression by antisense small RNA-mediated transcriptional gene silencing (gs) resulted in growth retardation, delayed endocytosis, and reduced acidification of endosomes and phagosomes. In this study, to better understand the role of Atg8 in phagocytosis and trogocytosis, we conducted a comparative proteomic analysis of phagosomes isolated from wild type and atg8-gs strains. We found that 127 and 107 proteins were detected >1.5-fold less or more abundantly, respectively, in phagosomes isolated from the atg8-gs strain, compared to the control strain. Among 127 proteins whose abundance was reduced in phagosomes from atg8-gs, a panel of proteins related to fatty acid metabolism, phagocytosis, and endoplasmic reticulum (ER) homeostasis was identified. Various lysosomal hydrolases and their receptors also tend to be excluded from phagosomes by atg8-gs, reinforcing the notion that Atg8 is involved in phagosomal acidification and digestion. On the contrary, among 107 proteins whose abundance increased in phagosomes from atg8-gs strain, ribosome-related proteins and metabolite interconversion enzymes are enriched. We further investigated the localization of several representative proteins, including adenylyl cyclase-associated protein and plasma membrane calcium pump, both of which were demonstrated to be recruited to phagosomes and trogosomes via an Atg8-dependent mechanism. Taken together, our study has provided the basis of the phagosome proteome to further elucidate molecular events in the Atg8-dependent regulatory network of phagosome/trogosome biogenesis in E. histolytica.
影响因子：
6.7
作者：
Boettner DR;Huston CD;Linford AS;Buss SN;Houpt E;Sherman NE;Petri WA Jr
通讯作者：
Petri WA Jr