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SRX4993761: GSM3463548: wild type, with 3-OH-C10-HSL, replicate 2; Methylomonas sp. LW13; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 37.3M spots, 11.2G bases, 4Gb downloads

Submitted by: NCBI (GEO)
Study: Interspecies chemical signalling in a methane-oxidizing bacterial community
show Abstracthide Abstract
Multiple species of bacteria oxidize methane in the environment after it is produced by anaerobic ecosystems. These organisms provide a carbon and energy source for species that cannot oxidize methane themselves, thereby serving a key role in these niches while also sequestering this potent greenhouse gas before it enters the atmosphere. Deciphering the molecular details of how methane-oxidizing bacteria interact in the environment enables us to understand an important aspect that shapes the structure and function these communities. Here we show that many members of the Methylomonas genus possess a LuxR-type acyl-homoserine lactone (acyl-HSL) receptor/transcription factor highly homologous to MbaR from the quorum sensing (QS) system of Methylobacter tundripaludum, another methane-oxidizer that has been isolated from the same environment. We reconstitute this detection system in Escherichia coli and also use mutant and transcriptomic analysis to show that the receptor from Methylomonas species strain LW13 (LW13) is active and alters LW13 gene expression in response to the acyl-HSL produced by M. tundripaludum. These findings provide a molecular mechanism for how two species of bacteria that may compete for resources in the environment can interact in a specific manner through a chemical signal. Overall design: In order to determine which genes MmsR regulates in response to acyl-HSL signal, we compared the transcriptome of exponentially growing LW13 in the presence or absence of 3-OH-C10-HSL. As a control for gene changes that are not the result of signal binding to MmsR, we constructed an unmarked, in-frame deletion of mmsR (?mmsR) and also compared the transcriptome of this strain in the presence and absence of signal. For each condition, we sequenced the transcriptomes of two biological replicates.
Sample: wild type, with 3-OH-C10-HSL, replicate 2
SAMN10393022 • SRS4029173 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Cell pellets were lysed by bead beating with 0.1-mm zirconia-silica beads (Biospec Products) in 1 mL TRIzol (Thermo Fisher Scientific). The lysate was then separated according to the TRIzol instructions, and subsequently 1.5 volumes of 100% ethanol was added to the aqueous phase of the extract which was then used as the input for an RNeasy RNA isolation kit (Qiagen). The resulting elution was then digested using Ambion DNase I (Thermo Fisher Scientific) for 30 minutes at 37ºC before being re-purified via RNeasy RNA isolation kit with the addition of an on-column DNase (Qiagen) digestion step for 15 minutes at room temperature. The resulting purified RNA was checked for DNA contamination by PCR using the universal 16S primers 27F and 1492R. cDNA library preparation and RNA sequencing were performed by GENEWIZ using Illumina HiSeq 2x150 (pair ended) reads.
Experiment attributes:
GEO Accession: GSM3463548
Links:
Runs: 1 run, 37.3M spots, 11.2G bases, 4Gb
Run# of Spots# of BasesSizePublished
SRR817320837,341,50311.2G4Gb2019-02-04

ID:
6727480

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