Dataset Information

Quantitative analysis of proteomic changes induced by potential quorum sensing in Haloferax volcanii

ABSTRACT: Quorum sensing (QS) is a mechanism of intercellular communication that enables microbes to alter gene expression and adapt to the environment. While QS has been extensively studied in bacteria, little is known about cell-cell communication in archaea. We could show that for Haloferax volcanii, the transition from motile rods to non-motile disks is dependent on a possibly novel, secreted small molecule present in cell-free conditioned medium (CM). Moreover, we determined that this putative QS molecule fails to induce the morphology transition in mutants lacking potential regulatory factors, DdfA and CirA. Quantitative proteomics of wild-type cells demonstrated that at least 236 proteins have significant differential abundances in the presence of CM. Conversely, in the ∆ddfA mutant, addition of CM resulted in only 110 proteins of significant differential abundances. These results confirm that DdfA is involved in CM-dependent regulation. CirA, along with other proteins involved in morphology and swimming motility transitions, is among the proteins regulated by DdfA. These discoveries significantly advance our understanding of microbial communication within archaeal species. Please note that this dataset is an extension to PXD040781. The raw files included here are additional MS measurements, but the search and quantification results correspond to a combination of all MS files (incl. those from PXD040781).

INSTRUMENT(S):

ORGANISM(S): Archaea Haloferax Volcanii (halobacterium Volcanii)

SUBMITTER: Stefan Schulze

LAB HEAD: Mechthild Pohlschroder

PROVIDER: PXD059278 | Pride | 2025-06-17

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20220622_jcl_SSchulze_Pohlschroder_UPenn_Haloferax_H53_3a_330ng_60min_Wat25cmBEH_1.raw	Raw
	20220622_jcl_SSchulze_Pohlschroder_UPenn_Haloferax_H53_3b_330ng_60min_Wat25cmBEH_1.raw	Raw
	20220622_jcl_SSchulze_Pohlschroder_UPenn_Haloferax_H53_3c_330ng_60min_Wat25cmBEH_1.raw	Raw
	20220622_jcl_SSchulze_Pohlschroder_UPenn_Haloferax_d2176_8a_330ng_60min_Wat25cmBEH_1.raw	Raw
	20220622_jcl_SSchulze_Pohlschroder_UPenn_Haloferax_d2176_8b_330ng_60min_Wat25cmBEH_1.raw	Raw

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Publications

Quorum sensing mediates morphology and motility transitions in the model archaeon <i>Haloferax volcanii</i>.

Chatterjee Priyanka P Consoli Caroline E CE Schiller Heather H Winter Kiersten K KK McCallum Monica E ME Schulze Stefan S Pohlschroder Mechthild M

bioRxiv : the preprint server for biology 20250114

Quorum sensing (QS) is a mechanism of intercellular communication that enables microbes to alter gene expression and adapt to the environment. This cell-cell signaling is necessary for intra- and interspecies behaviors such as virulence and biofilm formation. While QS has been extensively studied in bacteria, little is known about cell-cell communication in archaea. Here we established an archaeal model system to study QS. We showed that for <i>Haloferax volcanii,</i> the transition from motile ...[more]

PMID: 39868331

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Project description:The genus Lactobacillus contains over 100 different species that were traditionally considered to be uniformly non-motile. However, at least twelve motile species are known to exist in the L. salivarius clade of this genus. Of these, Lactobacillus rumnis is the only motile species that is also autochthonous to the mammalian gastrointestinal tract. The genomes of two L. ruminis strains, ATCC25644 (human isolate, non-motile) and ATCC27782 (bovine isolate, motile) were sequenced and annotated to identify the genes responsible for flagellum biogenesis and chemotaxis in this species. Transcriptome analysis revealed that motility genes were transcribed at a significantly higher level in motile L. ruminis ATCC27782 than in non-motile ATCC25644 during the motile growth phase. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the motile phase, 20 ml aliquots of MRS broth were inoculated (0.25 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 Â°C for 15 hr. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the non-motile growth phase, 20 ml aliquots of MRS broth were inoculated (1 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 Â°C for 16-20 hrs. RNAprotect Bacteria reagent (Qiagen) was used to stabilize gene expression in the target cultures, and total RNA was isolated according to the protocol for enzymatic and mechanical disruption of bacteria as described in the RNAprotect Bacteria Reagent handbook. RNA isolation was completed with the RNeasy Mini kit, and contaminating DNA was removed with the Turbo DNA-free kit (Ambion). An Oligo aCGH/ChIP-on chip hybridization kit (Agilent) was used for hybridization of the labelled cDNA to the microarrays. Probe hybridization took place at 65 Â°C for 20 hrs with constant rotation (10 rpm). Microarrays were scanned using the Agilent Microarray Scanner System (G2505B) and the scanned files were converted to data files with Feature Extraction software (Agilent). Three microarrays, (three biological replicates) were used to examine gene transcription during the motile growth phase. One microarray was used to examine gene transcription during the non-motile growth phase.

Project description:Quorum-sensing (QS) is a cell-cell communication system that controls gene expression in many bacterial species, mediated by diffusible signal molecules. While the intracellular regulatory mechanisms of QS are often well-understood, the functional roles of QS remain controversial. In particular, the use of multiple signals by many bacterial species poses a serious challenge to current functional theories. Here we address this challenge by showing that bacteria can use multiple QS signals to infer both their social (density) and physical (mass-transfer) environment. Analytical and evolutionary simulation models show that the detection of and response to complex social/physical contrasts requires multiple signals with distinct half-lives and combinatorial (non-additive) responses to signal concentrations. We test these predictions using the opportunistic pathogen Pseudomonas aeruginosa, and demonstrate significant differences in signal decay between its two primary signal molecules as well as diverse combinatorial responses to dual signal inputs. QS is associated with the control of secreted factors, and we show that secretome genes are preferentially controlled by synergistic M-bM-^@M-^XAND-gateM-bM-^@M-^Y responses to multiple signal inputs, ensuring the effective expression of secreted factors in high density and low mass-transfer environments. Our results support a novel functional hypothesis for the use of multiple signals and, more generally, show that bacteria are capable of combinatorial communication. The two primary signal molecules of P. aeruginosa are the homoserine lactones N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) and N-butyryl-homoserine lactone (C4-HSL). Effects of the different signal molecules was assessed using a double QS synthase mutant of Pseudomonas aeruginosa PAO1 lasI/rhlI grown at 37M-BM-0C in 25 ml LB broth and 250 ml flasks with shaking at 200 r.p.m. in four treatments, each with a replicate: (a) no addition; (b) 3-oxo- C12-HSL; (c) C4-HSL; and (d) both 3-oxo-C12-HSL and C4-HSL.

Dataset Information

Quantitative analysis of proteomic changes induced by potential quorum sensing in Haloferax volcanii

Dataset's files

Publications

Quorum sensing mediates morphology and motility transitions in the model archaeon <i>Haloferax volcanii</i>.

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Dataset Information

Quantitative analysis of proteomic changes induced by potential quorum sensing in Haloferax volcanii

Dataset's files

Publications

Quorum sensing mediates morphology and motility transitions in the model archaeon &lt;i&gt;Haloferax volcanii&lt;/i&gt;.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Quorum sensing mediates morphology and motility transitions in the model archaeon <i>Haloferax volcanii</i>.