1. The Burkholderia pseudomallei intracellular 'TRANSITome'.
- Author
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Heacock-Kang Y, McMillan IA, Norris MH, Sun Z, Zarzycki-Siek J, Bluhm AP, Cabanas D, Norton RE, Ketheesan N, Miller JF, Schweizer HP, and Hoang TT
- Subjects
- Animals, Burkholderia pseudomallei cytology, Burkholderia pseudomallei pathogenicity, Cell Line, Tumor, Cell Membrane microbiology, Cytoplasm microbiology, HEK293 Cells, Host-Pathogen Interactions, Humans, Melioidosis microbiology, Mice, Mice, Inbred BALB C, RAW 264.7 Cells, Single-Cell Analysis methods, Vacuoles microbiology, Virulence genetics, Bacterial Proteins genetics, Burkholderia pseudomallei genetics, Computational Biology methods, Gene Expression Profiling methods, Genes, Bacterial genetics, Virulence Factors genetics
- Abstract
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.
- Published
- 2021
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