Anton Leonie, Cheng Wenjing, Haile Meseret T, Dziekan Jerzy M, Cobb David W, Zhu Xiyan, Han Leyan, Li Emerson, Nair Anjali, Lee Carolyn L, Wang Hanyu, Ke Hangjun, Zhang Guoan, Doud Emma H, Cowman Alan F, Ho Chi-Min
Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, USA.
Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.
Nat Struct Mol Biol. 2025 Aug 18. doi: 10.1038/s41594-025-01632-3.
Our understanding of cellular events is hampered by the gap between the resolution at which we can observe events inside cells and our ability to replicate physiological conditions in test tubes. Here, we show in Plasmodium falciparum, a non-model organism of high medical importance, that this gap can be bridged by using an integrated structural biology approach to visualize events inside the cell at molecular resolution. We determined eight high-resolution structures of the native malarial ribosome in actively translating states inside P. falciparum-infected human erythrocytes using in situ cryo-electron tomography. Following perturbation with a Plasmodium-specific translation inhibitor, we then observed a decrease in elongation factor-bound ribosomal states and an apparent upregulation of ribosome biogenesis in inhibitor-treated parasites. Our work elucidates new molecular details of the malarial translation elongation cycle and demonstrates direct multiscale visualization of drug-induced phenotypic changes in the structure and localization of individual molecules within the native cellular context.
我们对细胞内事件的理解受到限制，因为我们观察细胞内事件的分辨率与在试管中复制生理条件的能力之间存在差距。在这里，我们在恶性疟原虫（一种具有高度医学重要性的非模式生物）中表明，通过使用综合结构生物学方法以分子分辨率可视化细胞内事件，可以弥合这一差距。我们使用原位冷冻电子断层扫描技术，确定了在恶性疟原虫感染的人类红细胞内处于活跃翻译状态的天然疟疾核糖体的八个高分辨率结构。在用疟原虫特异性翻译抑制剂进行扰动后，我们随后观察到在抑制剂处理的寄生虫中，延伸因子结合的核糖体状态减少，核糖体生物合成明显上调。我们的工作阐明了疟疾翻译延伸循环的新分子细节，并展示了在天然细胞环境中对药物诱导的单个分子结构和定位的表型变化进行直接多尺度可视化。
