Dataset Information

Archael ubiquitin-like SAMP3 is isopeptide-linked to Proteins by an UbaA-dependent mechanism

ABSTRACT: FLAG-SAMP3ylated proteins were immunoprecipitated from Haloferax volcanii using anti-FLAG affinity agarose and separated using SDS-PAGE. The Coomassie-stained IP-bands represented FLAG-SAMP3ylated proteins harvested from the wild type and also from the ubaA mutant as negative control. The Coomassie-lanes from the IP samples were cut into 10 fractions and in-gel tryptic digested and the tryptic peptides measured using Orbitrap Velos LC-MS/MS.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Haloferax Volcanii (halobacterium Volcanii)

SUBMITTER: Haike Antelmann

LAB HEAD: Haike Antelmann

PROVIDER: PXD000202 | Pride | 2020-07-02

REPOSITORIES: Pride

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Publications

The Archaeal Proteome Project advances knowledge about archaeal cell biology through comprehensive proteomics.

Schulze Stefan S Adams Zachary Z Cerletti Micaela M De Castro Rosana R Ferreira-Cerca Sébastien S Fufezan Christian C Giménez María Inés MI Hippler Michael M Jevtic Zivojin Z Knüppel Robert R Legerme Georgio G Lenz Christof C Marchfelder Anita A Maupin-Furlow Julie J Paggi Roberto A RA Pfeiffer Friedhelm F Poetsch Ansgar A Urlaub Henning H Pohlschroder Mechthild M

Nature communications 20200619 1

While many aspects of archaeal cell biology remain relatively unexplored, systems biology approaches like mass spectrometry (MS) based proteomics offer an opportunity for rapid advances. Unfortunately, the enormous amount of MS data generated often remains incompletely analyzed due to a lack of sophisticated bioinformatic tools and field-specific biological expertise for data interpretation. Here we present the initiation of the Archaeal Proteome Project (ArcPP), a community-based effort to comp ...[more]

PMID: 32561711

Publication: 1/2

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Project description:To cause a systemic infection, Salmonella must respond to many environmental cues during mouse infection and express specific subsets of genes in a temporal and spatial manner, but the regulatory pathways are poorly established. To unravel how micro-environmental signals are processed and integrated into coordinated action, we constructed in-frame non-polar deletions of 83 regulators inferred to play a role in Salmonella enteriditis Typhimurium (STM) virulence and tested them in three virulence assays (intraperitoneal [i.p.], and intragastric [i.g.] infection in BALB/c mice, and persistence in 129X1/SvJ mice). Overall, 35 regulators were identified whose absence attenuated virulence in at least one assay, and of those, 14 regulators were required for systemic mouse infection, the most stringent virulence assay. As a first step towards understanding the interplay between a pathogen and its host from a systems biology standpoint, we focused on these 14 genes. Transcriptional profiles were obtained for deletions of each of these 14 regulators grown under four different environmental conditions. These results, as well as publicly available transcriptional profiles, were analyzed using both network inference and cluster analysis algorithms. The analysis predicts a regulatory network in which all 14 regulators control the same set of genes necessary for Salmonella to cause systemic infection. We tested the regulatory model by expressing a subset of the regulators in trans and monitoring transcription of 7 known virulence factors located within Salmonella pathogenicity island 2 (SPI-2). These experiments validated the regulatory model and showed that the response regulator SsrB and the MarR type regulator, SlyA, are the terminal regulators in a cascade that integrates multiple signals. Furthermore, experiments to demonstrate epistatic relationships showed that SsrB can replace SlyA and, in some cases, SlyA can replace SsrB for expression of SPI-2 encoded virulence factors. A total of 55 RNA pools from 14 strains in 4 conditions were prepared. An additional 12 RNA samples were prepared from the parental strain. All were subjected to microarray analysis according to the procedures described (Porwollik S et al. 2003 Nucleic Acids Res 31: 1869M-bM-^@M-^S1876). Genomic DNA from 14028s was isolated and used as reference for hybridization.

Dataset Information

Archael ubiquitin-like SAMP3 is isopeptide-linked to Proteins by an UbaA-dependent mechanism

Publications

The Archaeal Proteome Project advances knowledge about archaeal cell biology through comprehensive proteomics.

Similar Datasets

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