School of Life Sciences, University of Hawai'i at Mānoa, Honolulu, HI, USA.
Department of Geography and Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA.
Nat Commun. 2021 Mar 26;12(1):1907. doi: 10.1038/s41467-021-22169-1.
Prokaryotic cell transcriptomics has been limited to mixed or sub-population dynamics and individual cells within heterogeneous populations, which has hampered further understanding of spatiotemporal and stage-specific processes of prokaryotic cells within complex environments. Here we develop a 'TRANSITomic' approach to profile transcriptomes of single Burkholderia pseudomallei cells as they transit through host cell infection at defined stages, yielding pathophysiological insights. We find that B. pseudomallei transits through host cells during infection in three observable stages: vacuole entry; cytoplasmic escape and replication; and membrane protrusion, promoting cell-to-cell spread. The B. pseudomallei 'TRANSITome' reveals dynamic gene-expression flux during transit in host cells and identifies genes that are required for pathogenesis. We find several hypothetical proteins and assign them to virulence mechanisms, including attachment, cytoskeletal modulation, and autophagy evasion. The B. pseudomallei 'TRANSITome' provides prokaryotic single-cell transcriptomics information enabling high-resolution understanding of host-pathogen interactions.
原核细胞转录组学一直受到混合或亚群动态以及异质群体中单个细胞的限制，这阻碍了对复杂环境中原核细胞时空和特定阶段特定过程的进一步理解。在这里，我们开发了一种“TRANSITomic”方法，以描绘单个伯克霍尔德氏菌假单胞菌细胞在宿主细胞感染过程中经过定义阶段时的转录组，从而获得病理生理学见解。我们发现，伯克霍尔德氏菌假单胞菌在感染过程中通过宿主细胞经历三个可观察到的阶段：液泡进入；细胞质逃逸和复制；以及膜突出，促进细胞间传播。伯克霍尔德氏菌假单胞菌的“TRANSITome”揭示了宿主细胞中转运过程中的动态基因表达通量，并确定了致病所必需的基因。我们发现了几个假设蛋白，并将它们分配到毒力机制中，包括附着、细胞骨架调节和自噬逃避。伯克霍尔德氏菌假单胞菌的“TRANSITome”提供了原核单细胞转录组学信息，能够实现对宿主-病原体相互作用的高分辨率理解。
