吞噬作用,细胞吞噬大颗粒的过程,在驱动组织清除和宿主防御中起着至关重要的作用。它的失调与自身免疫有关,蛋白质的毒性积累,和增加感染的风险。尽管它很重要,我们对这个过程中涉及的所有分子成分缺乏充分的了解。为了在人类细胞中创建功能图,我们进行了全基因组CRISPRkoFACS筛选,鉴定出716个基因.将这些命中映射到针对功能性细胞过程注释的全面的蛋白质-蛋白质相互作用网络,允许多次鉴定的蛋白质复合物的检索和缺失的吞噬作用调节因子的检测。除了已知的组件,例如Arp2/3复合物,液泡-ATPase-Rag机制,和Wave-2复合体,我们确定并验证了新的吞噬作用相关功能,包括寡糖转移酶复合物(MAGT1/SLC58A1,DDOST,STT3B,和RPN2)和hypusine途径(eIF5A,DHPS,和DOHH)。总的来说,我们的吞噬作用网络包括货物摄取的元素,洗牌,通过细胞进行生物转化,为鉴定内溶酶体系统疾病的潜在新驱动因素提供了资源。我们整合蛋白质-蛋白质相互作用的方法为功能上解释全基因组筛选提供了广泛适用的方法。
Phagocytosis, the process by which cells engulf large particles, plays a vital role in driving tissue clearance and host defense. Its dysregulation is connected to autoimmunity, toxic accumulation of proteins, and increased risks for infections. Despite its importance, we lack full understanding of all molecular components involved in the process. To create a functional map in human cells, we performed a genome-wide CRISPRko FACS screen that identified 716 genes. Mapping those hits to a comprehensive protein-protein interaction network annotated for functional cellular processes allowed retrieval of protein complexes identified multiple times and detection of missing phagocytosis regulators. In addition to known components, such as the Arp2/3 complex, the vacuolar-ATPase-Rag machinery, and the Wave-2 complex, we identified and validated new phagocytosis-relevant functions, including the oligosaccharyltransferase complex (MAGT1/SLC58A1, DDOST, STT3B, and RPN2) and the hypusine pathway (eIF5A, DHPS, and DOHH). Overall, our phagocytosis network comprises elements of cargo uptake, shuffling, and biotransformation through the cell, providing a resource for the identification of potential novel drivers for diseases of the endo-lysosomal system. Our approach of integrating protein-protein interaction offers a broadly applicable way to functionally interpret genome-wide screens.