Control of type III protein secretion using a minimal genetic system.

Nat Commun
Authors
Abstract

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.

Year of Publication
2017
Journal
Nat Commun
Volume
8
Pages
14737
Date Published
2017 May 09
ISSN
2041-1723
DOI
10.1038/ncomms14737
PubMed ID
28485369
PubMed Central ID
PMC5436071
Links
Grant list
P50 GM098792 / GM / NIGMS NIH HHS / United States
R01 AI067699 / AI / NIAID NIH HHS / United States