Dataset Information

Haloferax volcanii_Shotgun_Archaeal Proteome Project

ABSTRACT: Archaea are ubiquitous prokaryotes with a wide range of habitats, important roles in ecology, biotechnology and potentially even human health. Despite that, our understanding of archaeal cell biology is still rather limited, partially because the application of systems biology approaches is lacking behind the other domains of life. Here we introduce/announce the Archaeal Proteome Project (ArcPP), a community effort that aims for the comprehensive analysis of archaeal proteomes. Starting with the model archaeon Haloferax volcanii, we have re-analyzed more than 2 TB of MS result files (>20 Mio. spectra) using state-of-the-art bioinformatic tools, increasing peptide spectrum matches and leading to the secure identification of >3000 proteins. This dataset is part of the Archaeal Proteome Project dataset

INSTRUMENT(S): Q Exactive

ORGANISM(S): Haloferax Volcanii Ds2

TISSUE(S): Cell Line Cell

SUBMITTER: Sébastien Ferreira-Cerca

LAB HEAD: Sébastien Ferreira-Cerca

PROVIDER: PXD014974 | Pride | 2020-05-11

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	MaxQuant_Output_txt.zip	Other
	POL11409X002__A_1_92-59212.raw	Raw
	POL11409X003__A_2_92-59213.raw	Raw

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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

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Project description:In this study the transcriptome of Haloferax volcanii DS70 (Wendoloski D, Ferrer C, Dyall-Smith ML. Microbiology. 2001, 147: 959-64), grown under of a variety of conditions, was mapped using a custom genome-wide tiled array consisting of 25 nt probes spaced in 35 nt windows. H. volcanii is a facultative aerobe capable of growth on simple carbon and nitrogen sources, it is amenable to genetic manipulations and has become a widely used model organism for studies on haloarchaeal physiology and global gene regulation. Of particular interest is the identification of genes encoding enzymes needed for growth on simple carbon and nitrogen sources and how these cells respond to limitations in these key nutrients. To address these questions we have compared the RNA populations of cells undergoing balanced growth in completely defined minimal medium (CDM) with succinate-glycerol, pyruvate or lactate as sole carbon sources with RNA from cells grown in complex rich medium (CX) containing yeast extract and tryptone. Data from these studies have identified a core group of genes expressing during balanced growth in either condition as well as specific genes associated with the biosynthesis of amino acids, nucleotides and other key building blocks. The response to nutrient limitation was also investigated. RNA populations from cells entering stationary phase from growth in CDM and CX media were compared to those undergoing balanced growth, and these RNAs were also compared with RNA populations of cells subjected to nitrogen or carbon starvation. Growth in the presence of histidine as the sole source of nitrogen in CDM medium also presented a model for growth with a “poor” nitrogen source. The results of these studies identified specific subpopulations of genes associated with nitrogen limitation and general nutrient limitation associated with the transition to stationary phase. Many of the responsive protein encoding genes were assigned general functions based on their similarity to known proteins; however, in each comparison, approximately half of the affected genes encode proteins classified as unknown or hypothetical proteins. The haloarchaea are also distinct among the Archaea in having multiple genes encoding the general transcription factor proteins TATA binding protein (TBP) and transcription factor IIB (TFB). We observed differential expression of these genes, providing further support for the proposal that alternative TBP-TFB pairings are associated with programmed changes in gene expression in the haloarchaea. RNA was prepared from H. volcanii DS70 cells grown under a variety of conditions. These include: balanced growth in completely defined medium with succinate and glycerol, pyruvate, or lactate as sole carbon source; balanced growth in complex medium with yeast extract and tryptone; starvation for nitrogen or carbon by re-suspending cells, undergoing balanced growth, in media lacking these agents; balanced growth in the presence of low (10% w/v NaCl) and high (20 % or 25% w/v NaCl) salt, and cells entering stationary from growth in CDM or CX media. Four independent cultures were prepared for each growth variable and RNA was isolated from each culture. RNA samples were identified as acceptable for use in array analysis if clear 23S rRNA, 16S rRNA and tRNA-sized bands were present in agarose gel and capillary electrophoresis analysis of the samples. Three RNA samples were chosen for each growth variable and each was used as the source material for a separate chip experiment; these constituted three biological replicates for the growth variable. Data from 48 chip hybridizations, representing 16 growth conditions, were combined for the quantile scaling (Auer H, Newsom DL, Nowak NJ, McHugh KM, Singh S, Yu CY, Yang Y, Wenger GD, Gastier-Foster JM, Kornacker K. BMC Genomics. 2007, 8:111-124) presented in the data analyses. Two additional experiments are included where the RNAs of three independent biological samples were pooled into a single RNA population and used as the source material for an array experiment. These included RNA samples enriched for species less than 500 nt that were isolated from cells undergoing balanced growing, and entry into stationary phase, in CDM or CX media. A second pooled series were obtained from the H. volcanii DS70 mutant strains ASD40 (∆pyrF) and ASD41 (∆pyrF∆hutR) grown in CDM with ammonium ion or histidine as the sole source of nitrogen. Pooled samples are presented as quantile normalized values (Bolstad, B.M., Irizarry, R.A., Astrand, M. and Speed, T.P. Bioinformatics. 2003, 19: 185-193). Total genomic DNA from H. volcanii DS70 was also used as probe, in two independent array experiments, to measure the inherent hybridization to the probes. These data are presented as quantile scaled values (Auer et. al., 2007).