PXD027465

PXD027465 is an original dataset announced via ProteomeXchange.

Title	The consequences of reciprocally exchanging the genomic sites of Integration Host Factor (IHF) subunit production for subunit stoichiometry and bacterial physiology in Salmonella enterica serovar Typhimurium
Description	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 that encode the subunits are located 350 kilobase pairs apart, in the Right replichore of the Salmonella chromosome. IHF is composed of one IhfA and one IhfB subunit. Despite this 1:1 stoichiometry, mass spectrometry revealed that IhfB is produced in 2-fold excess over IhfA. Results from several studies suggest that a gene's location on the bacterial chromosome is an important factor in establishing the appropriate spatiotemporal expression pattern of that gene and its influence on physiology. 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. Mass spectrometry 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, had identical cell morphology, and was similar in competitive fitness. However, it was less motile than the wild type, 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 that accompanies altered phenotypes.
HostingRepository	PRIDE
AnnounceDate	2021-11-02
AnnouncementXML	Submission_2021-11-02_03:35:34.825.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD027465
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	German Pozdeev
SpeciesList	scientific name: Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344; NCBI TaxID: 216597;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-07-21 05:41:18	ID requested
⏵ 1	2021-11-02 03:35:35	announced

Dataset with its publication pending

submitter keyword: Salmonella enterica serovar Typhimurium

Integration Host Factor

IHF

nucleoid-associated protein

LC-MSMS

stoichiometry

Charles James Dorman
contact affiliation	Trinity College Dublin
contact email	cjdorman@tcd.ie
lab head
German Pozdeev
contact affiliation	Royal College of Surgeons in Ireland
contact email	germanpozdeev@rcsi.ie
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD027465
     1. Label: PRIDE project
     2. Name: The consequences of reciprocally exchanging the genomic sites of Integration Host Factor (IHF) subunit production for subunit stoichiometry and bacterial physiology in Salmonella enterica serovar Typhimurium