Temporal profiling of Salmonella transcriptional dynamics during macrophage infection using a comprehensive reporter library
Abstract/Contents
- Abstract
- The transcriptome of Salmonella enterica serovar Typhimurium (S. Tm) dynamically responds to rapid environmental shifts intrinsic to S. Tm lifestyle, exemplified by entry into the Salmonella-containing vacuole (SCV) within macrophages. Intracellular S. Tm must respond to the acidity of the SCV, accumulation of reactive oxygen/nitrogen species, and fluctuations in nutrient availability. Despite thorough RNA-seq-based investigations, the precise transcriptional timing of the expression of many secretion systems, metabolic pathways, and virulence effectors involved in infection has yet to be elucidated. Here, we construct a comprehensive library of GFP-reporter strains representing ~3,000 computationally identified S. Tm promoter regions to study the dynamics of transcriptional regulation. We quantified promoter activity during in vitro growth in defined and complex media and throughout the timeline of intracellular infection of RAW 264.7 macrophages. Using bulk measurements and single-cell imaging, we uncovered condition-specific transcriptional regulation and population-level heterogeneity in the activity of SPI-2-related promoters. We discovered previously unidentified transcriptional activity from 234 promoters, including ones with novel activity during macrophage infection that are associated with pathogenicity islands and are involved in metal homeostasis and carbon metabolism, specifically the Entner-Doudoroff pathway. Our library and datasets provide powerful resources for systems-level interrogation of Salmonella transcriptional dynamics.
Description
| Type of resource | Dataset, text |
|---|---|
| Date modified | July 21, 2023; September 13, 2023; September 13, 2023; June 17, 2024 |
| Publication date | July 20, 2023 |
Creators/Contributors
| Author | Nguyen, Taylor |
https://orcid.org/0000-0002-1288-8882
(unverified)
|
|---|---|---|
| Author | Diaz, Oscar | |
| Author | Wang, Benjamin X. | |
| Research team head | Monack, Denise | |
| Research team head | Huang, Kerwyn C. |
Subjects
| Subject | transcriptional reporter fusion |
|---|---|
| Subject | transcriptomics |
| Subject | in vitro screening |
| Subject | macrophages |
| Subject | SPI2 |
| Subject | Entner-Doudoroff pathway |
| Subject | population-level heterogeneity |
| Subject | manganese |
| Subject | metal homeostasis |
| Genre | Data |
| Genre | Manuscript |
| Genre | Data sets |
| Genre | Dataset |
Bibliographic information
Access conditions
- Use and reproduction
- User agrees that, where applicable, content will not be used to identify or to otherwise infringe the privacy or confidentiality rights of individuals. Content distributed via the Stanford Digital Repository may be subject to additional license and use restrictions applied by the depositor.
- License
- This work is licensed under a Creative Commons Attribution 4.0 International license (CC BY).
Preferred citation
- Preferred citation
- Nguyen, T., Wang, B., Diaz, O., Monack, D., and Huang, K. (2023). Temporal profiling of Salmonella transcriptional dynamics during macrophage infection using a comprehensive reporter library. Stanford Digital Repository. Available at https://purl.stanford.edu/fc012fq3845. https://doi.org/10.25740/fc012fq3845.
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Stanford Research Data
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