Reciprocally rewiring and repositioning the Integration Host Factor (IHF) subunit genes in Salmonella enterica serovar Typhimurium: impacts on physiology and virulence. Issue 2 (15th February 2022)

• Record Type: Journal Article
• Authors: Pozdeev, German; Beckett, Michael C.; Mogre, Aalap; Thomson, Nicholas R.; Dorman, Charles J.
• Is Part Of: Microbial genomics. Volume 8:Issue 2(2022)
• Publication Date: 2022-02-15
• Subjects: Salmonella enterica serovar Typhimurium -- Integration Host Factor (IHF) -- genome organization -- genome rewiring -- proteome -- gene regulation; Microbial genomics -- Periodicals; 572.8629
• DOI: 10.1099/mgen.0.000768
• Languages: English
• ISSNs: 2057-5858

Abstract

Abstract : The Integration Host Factor (IHF) is a heterodimeric nucleoid-associated protein that plays roles in bacterial nucleoid architecture and genome-wide gene regulation. The ihfA and ihfB genes encode the subunits and are located 350 kbp apart, in the Right replichore of the Salmonella chromosome. IHF is composed of one IhfA and one IhfB subunit. Despite this 1 : 1 stoichiometry, MS revealed that IhfB is produced in 2-fold excess over IhfA. We re-engineered Salmonella to exchange reciprocally the protein-coding regions of ihfA and ihfB, such that each relocated protein-encoding region was driven by the expression signals of the other's gene. MS showed that in this 'rewired' strain, IhfA is produced in excess over IhfB, correlating with enhanced stability of the hybrid ihfB–ihfA mRNA that was expressed from the ihfB promoter. Nevertheless, the rewired strain grew at a similar rate to the wild-type and was similar in competitive fitness. However, compared to the wild-type, it was less motile, had growth-phase-specific reductions in SPI-1 and SPI-2 gene expression, and was engulfed at a higher rate by RAW macrophage. Our data show that while exchanging the physical locations of its ihf genes and the rewiring of their regulatory circuitry are well tolerated in Salmonella, genes involved in the production of type 3 secretion systems exhibit dysregulation accompanied by altered phenotypes.