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| Status |
Public on Oct 10, 2008 |
| Title |
Control of RpoS in global gene expression of Escherichia coli in minimal media |
| Organism |
Escherichia coli K-12 |
| Experiment type |
Expression profiling by array
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| Summary |
RpoS, an alternative sigma factor, is critical for stress response in Escherichia coli. The RpoS regulon expression has been well characterized in rich media that support fast growth and high growth yields. In contrast, though RpoS levels are high in minimal media, how RpoS functions under such conditions has not been clearly resolved. In this study, we compared the global transcriptional profiles of wild type and an rpoS mutant of E. coli grown in glucose minimal media using microarray analyses. The expression of over 200 genes was altered by loss of RpoS in exponential and stationary phases, with only 48 genes common to both conditions. The nature of the RpoS-controlled regulon in minimal media was substantially different from that expressed in rich media. Specifically, the expression of many genes encoding regulatory factors (e.g., hfq, csrA and rpoE) and genes in metabolic pathways (e.g., lysA, lysC and hisD) were regulated by RpoS in minimal media. In early exponential phase, protein levels of RpoS in minimal media were much higher than that in LB media, which may at least partly account for the observed difference in the expression of RpoS-controlled genes. Expression of genes required for flagellar function and chemotaxis was elevated in the rpoS mutant. Western blot analyses show that the flagella sigma factor FliA was expressed much higher in rpoS mutants than in WT in all phase of growth. Consistent with this, the motility of rpoS mutants was enhanced relative to WT. In conclusion, RpoS and its controlled regulators form a complex regulatory network that mediates the expression of a large regulon in minimal media.
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| Overall design |
A precise rpoS deletion mutant of MG1655 was constructed using the red recombinase method. Wild type and rpoS mutants were inoculated in triplicate into M63 glucose (0.2%) minimal media at a starting OD of 0.0001 and grown aerobically at 37oC. Cultures were harvested at OD600= 0.3 in exponential phase and at OD600= 1.5 in stationary phase. For RNA extraction, cultures were mixed directly with a boiling lysis buffer containing SDS and EDTA followed by acidic hot phenol (65C) to minimize RNA degradation. RNA samples were hybridized to Affymetrix E. coli Antisense Genome Array according to Affymetrix's standard protocols.
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| Contributor(s) |
Dong T, Schellhorn HE |
| Citation(s) |
18843507 |
| Submission date |
Sep 16, 2008 |
| Last update date |
Sep 20, 2012 |
| Contact name |
Herb E Schellhorn |
| E-mail(s) |
schell@mcmaster.ca
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| Phone |
9055259140-27316
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| Organization name |
McMaster University
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| Department |
Biology
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| Lab |
405
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| Street address |
1280 Main St. W.,
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| City |
Hamilton |
| State/province |
ON |
| ZIP/Postal code |
L8S4K1 |
| Country |
Canada |
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| Platforms (1) |
| GPL199 |
[Ecoli_ASv2] Affymetrix E. coli Antisense Genome Array |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA110989 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE12797_RAW.tar |
19.1 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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