Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

RNA-Seq analysis of quorum sensing-dependent genes in Burkholderia glumae BGR1

ABSTRACT: Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, whose expression is regulated by quorum sensing (QS). The QS systems of the bacterium rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate toxoflavin biosynthesis genes and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNA-Seq of the wild type Burkholderia glumae BGR1 and two QS-defective mutants, BGS2 (BGR1 tofI::Ω) and BGS9 (BGR1 qsmR::Ω), with two different types of culture conditions including 6hr liquid culture (before onset QS) and 10hr liquid culture (after onset QS). 3 samples examined: Burkholderia glumae BGR1 wild type, and two QS-defective mutants, BGS2 (BGR1 tofI::Ω) and BGS9 (BGR1 qsmR::Ω). Two conditions: cultured in LB media for 6hrs or 10hrs.

ORGANISM(S): Burkholderia glumae

SUBMITTER: Ingyu Hwang

PROVIDER: E-GEOD-36485 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Bacterial quorum sensing, cooperativity, and anticipation of stationary-phase stress.

Goo Eunhye E Majerczyk Charlotte D CD An Jae Hyung JH Chandler Josephine R JR Seo Young-Su YS Ham Hyeonheui H Lim Jae Yun JY Kim Hongsup H Lee Bongsoo B Jang Moon Sun MS Greenberg E Peter EP Hwang Ingyu I

Proceedings of the National Academy of Sciences of the United States of America 20121112 48

Acyl-homoserine lactone-mediated quorum sensing (QS) regulates diverse activities in many species of Proteobacteria. QS-controlled genes commonly code for production of secreted or excreted public goods. The acyl-homoserine lactones are synthesized by members of the LuxI signal synthase family and are detected by cognate members of the LuxR family of transcriptional regulators. QS affords a means of population density-dependent gene regulation. Control of public goods via QS provides a fitness b ...[more]

PMID: 23150539

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Project description:The two major methods for growing C. elegans are liquid and plate culture. Differences in oxygenation and other environmental variables could affect many experimentally relevant biological processes, such as metabolism, cellular structure, and gene regulation. Worms on solid support move with deliberate sinusoidal movements, while worms in liquid thrash rapidly and almost constantly, often appearing elongated. We investigated differential gene expression under these two commonly used growth conditions. Embryos were isolated from adults grown on plates. After hatching, animals were placed either on plates or in liquid under standard conditions, and grown until young adulthood, approximately 48 hours later. RNA was isolated independently from each growth, differentially labeled with a fluorescent dye, and mixed directly for comparative hybridization to NimbleGen DNA microarrays, which contained probes designed to measure expression from all predicted C. elegans coding genes. The experiment was performed in triplicate. Analysis of the expression data yielded surprisingly few genes with differential expression, about 1%. Overall, 176 genes were downregulated significantly (p<0.01; Student's t) in liquid culture relative to plates, but of these only 102 genes were down two-fold or more. A total of 121 genes were upregulated (p<0.01. Student's t) in liquid; of these, RNA corresponding to 50 increased by at least two-fold. Surprisingly few genes directly associated with a stress response were differentially regulated in this experiment. However, the single gene most strongly upregulated in liquid is nnt-1, which encodes a mitochondrial NADP transhydrogenase, which protects against oxidative stress (Arkbladt et al., 2005 PMID: 15890626). Other molecules associated with reducing oxidation are also upregulated in liquid culture, including thioredoxin, and two glutathione-S-transferases. As a class, however, the group of genes most differentially regulated encode collagens. A total of 12 collagen genes were downregulated in liquid culture relative to plates, while none are upregulated. Other genes affected include those encoding proteins involved in ubiquitin-mediated proteolysis, Ras signaling, transcriptional regulation, ribosome function, and multiple membrane-associated proteins. The limited changes in relative RNA levels under different culture conditions provide increased confidence in comparisons made between worms grown in liquid and solid culture, and more generally between datasets obtained in different labs under slightly different culture conditions. Knowing which genes are most responsive to different culture conditions inform us about how environment affects the animal, and should be of interest for comparison to other stress and longevity global datasets.

Dataset Information

RNA-Seq analysis of quorum sensing-dependent genes in Burkholderia glumae BGR1

Publications

Bacterial quorum sensing, cooperativity, and anticipation of stationary-phase stress.

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