 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Mar 02, 2023 |
| Title |
SigB modulates expression of novel SigB regulon members via Bc1009 in non-stressed and heat-stressed cells revealing its alternative roles in Bacillus cereus |
| Organism |
Bacillus cereus ATCC 14579 |
| Experiment type |
Expression profiling by array
|
| Summary |
Background: The Bacillus cereus Sigma B (SigB) dependent general stress response is activated via the two-component RsbKY system, which involves a phosphate transfer from RsbK to RsbY. It has been hypothesized that the Hpr-like phosphocarrier protein (Bc1009) encoded by bc1009 in the SigB gene cluster may play a role in this transfer, thereby acting as a regulator of SigB activation. Alternatively, Bc1009 may be involved in the activation of a subset of SigB regulon members. Results: We first investigated the potential role of bc1009 to act as a SigB regulator but ruled out this possibility as the deletion of bc1009 did not affect the expression of sigB and other SigB gene cluster members. The SigB-dependent functions of Bc1009 were further examined in B. cereus ATCC14579 via comparative proteome profiling (backed up by transcriptomics) of wt, Δbc1009 and ΔsigB deletion mutants under heat stress at 42°C. This revealed 284 proteins displaying SigB-dependent alterations in protein expression levels in heat-stressed cells, including a subgroup of 138 proteins for which alterations were also Bc1009-dependent. Next to proteins with roles in stress defense, newly identified SigB and Bc1009-dependent proteins have roles in cell motility, signal transduction, transcription, cell wall biogenesis, and amino acid transport and metabolism. Analysis of lethal stress survival at 50°C after pre-adaptation at 42°C showed intermediate survival efficacy of Δbc1009 cells, highest survival of wt, and lowest survival of ΔsigB cells, respectively. Additional comparative proteome analysis of non-stressed wt and mutant cells at 30°C revealed 96 proteins with SigB and Bc1009-dependent differences in levels, 51 were also identified under heat stress, and 45 showed significant differential expression at 30°C, including proteins with roles in carbohydrate/ion transport and metabolism. Overlapping functions at 30°C and 42°C included proteins involved in motility, and ΔsigB and Δbc1009 cells showed reduced motility compared to wt cells in swimming assays at both temperatures. Conclusion: our results extend the B. cereus SigB regulon to >300 members, with a novel role of SigB-dependent Bc1009 in the activation of a subregulon of >180 members, conceivably via phosphotransfer-based interactions with other transcriptional regulatory networks.
|
| |
|
| Overall design |
Seven comparisons were performed: WT over ΔsigB at 30°C and 42°C, WT over Δbc1009 at 30°C and 42°C, WT 30°C over WT 42°C, ΔsigB 30°C over ΔsigB 42°C, Δbc1009 30°C over Δbc1009 42°C (all biological duplicate, and Cy5-Cy3 dye-swapped). Strains were grown in BHI at 30 degrees C until an OD600nm ~0.4 followed by exposure in the same media at 30 degrees C or 42 degrees C, while shaken at 160 rpm, and harvested after 20 minutes.
|
| |
|
| Contributor(s) |
Yeak KY, Tempelaars MH, Wu JL, Westerveld W, Reder A, Michalik S, Dhople VM, Völker U, Pané-Farré J, Wells-Bennik M, Abee T |
| Citation(s) |
36759782 |
| |
| Submission date |
Sep 22, 2022 |
| Last update date |
Mar 03, 2023 |
| Contact name |
Jeroen Steven Koomen |
| E-mail(s) |
Jeroen.Koomen@wur.nl
|
| Organization name |
Wageningen University
|
| Lab |
Food Microbiology
|
| Street address |
Bornse Weilanden 9
|
| City |
Wageningen |
| ZIP/Postal code |
6708 WG |
| Country |
Netherlands |
| |
|
| Platforms (1) |
| GPL9493 |
Bacillus cereus ATCC 14579, 3rd design, Agilent 8X15K |
|
| Samples (16)
|
|
| Relations |
| BioProject |
PRJNA883372 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE213958_RAW.tar |
17.2 Mb |
(http)(custom) |
TAR (of TXT) |
| GSE213958_ratios.txt.gz |
517.0 Kb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data are available on Series record |
| Processed data provided as supplementary file |
|
|
|
|
 |
Less...
More...