Nguyen, Taylor H 
ORCID: 0000-0002-1288-8882, Diaz, Oscar R, Rajendram, Manohary, Butler, Daniel SC, Wang, Benjamin X, Hinton, Jay CD ORCID: 0000-0003-2671-6026, Monack, Denise and Huang, Kerwyn Casey
(2023)
Temporal profiling of<i>Salmonella</i>transcriptional dynamics during macrophage infection using a comprehensive reporter library.
[Preprint]
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Nguyen etal 2023 Temporal profiling of Salmonella transcriptional dynamics in macrophages - BioRxiv.pdf - Preprint version Download (7MB) | Preview |
Abstract
<jats:title>Abstract</jats:title><jats:p>The transcriptome of<jats:italic>Salmonella enterica</jats:italic>serovar Typhimurium (<jats:italic>S</jats:italic>. Tm) dynamically responds to the rapid environmental shifts intrinsic to<jats:italic>S.</jats:italic>Tm lifestyle, exemplified by entry into the<jats:italic>Salmonella</jats:italic>-containing vacuole (SCV) within macrophages. Intracellular<jats:italic>S</jats:italic>. 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<jats:italic>S.</jats:italic>Tm promoter regions to study the dynamics of transcriptional regulation. We quantified promoter activity during<jats:italic>in vitro</jats:italic>growth in defined and complex media and throughout the timeline of intracellular infection of RAW 246.7 macrophages. Using bulk measurements and single-cell imaging, we uncovered condition-specific transcriptional regulation and population-level heterogeneity in the activity of virulence-related promoters, including SPI2 genes such as<jats:italic>ssaR</jats:italic>and<jats:italic>ssaG</jats:italic>. We discovered previously unidentified transcriptional activity from 234 genes, including ones with novel activity during infection that are associated with pathogenecity islands and are involved in metabolism and metal homeostasis. Our library and data sets should provide powerful resources for systems-level interrogation of<jats:italic>Salmonella</jats:italic>transcriptional dynamics.</jats:p>
| Item Type: | Preprint |
|---|---|
| Uncontrolled Keywords: | Vaccine Related, Genetics, Emerging Infectious Diseases, Infectious Diseases, Foodborne Illness, Prevention, 2.2 Factors relating to the physical environment, 2 Aetiology, 2.1 Biological and endogenous factors, Infection |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 07 Nov 2023 10:33 |
| Last Modified: | 25 Apr 2024 22:39 |
| DOI: | 10.1101/2023.09.27.559620 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3176666 |
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