Control of type III protein secretion using a minimal genetic system.
Miryoung SongDavid J SukovichLuciano CiccarelliJulia MayrJesus Fernandez-RodriguezEthan A MirskyAlex C TuckerD Benjamin GordonThomas C MarlovitsChristopher A VoigtPublished in: Nature communications (2017)
Gram-negative bacteria secrete proteins using a type III secretion system (T3SS), which functions as a needle-like molecular machine. The many proteins involved in T3SS construction are tightly regulated due to its role in pathogenesis and motility. Here, starting with the 35 kb Salmonella pathogenicity island 1 (SPI-1), we eliminated internal regulation and simplified the genetics by removing or recoding genes, scrambling gene order and replacing all non-coding DNA with synthetic genetic parts. This process results in a 16 kb cluster that shares no sequence identity, regulation or organizational principles with SPI-1. Building this simplified system led to the discovery of essential roles for an internal start site (SpaO) and small RNA (InvR). Further, it can be controlled using synthetic regulatory circuits, including under SPI-1 repressing conditions. This work reveals an incredible post-transcriptional robustness in T3SS assembly and aids its control as a tool in biotechnology.
Keyphrases
- type iii
- genome wide
- transcription factor
- copy number
- genome wide identification
- dna methylation
- biofilm formation
- escherichia coli
- gene expression
- single molecule
- small molecule
- amino acid
- deep learning
- nucleic acid
- ultrasound guided
- high throughput
- genome wide analysis
- antiretroviral therapy
- pseudomonas aeruginosa
- binding protein
- protein protein
- single cell
- candida albicans
- heat shock