 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Apr 21, 2020 |
| Title |
The DEAD-box RNA helicase CshA is required for fatty acid homeostasis in Staphylococcus aureus |
| Organism |
Staphylococcus aureus |
| Experiment type |
Other
|
| Summary |
Staphylococcus aureus is an opportunistic pathogen that can grow in wide array of conditions: on abiotic surfaces, on the skin, in the nose, in planktonic or biofilms forms and can cause many type of infections. Consequently, S. aureus must be able to adapt rapidly to these changing growth conditions, an ability largely driven at the posttranscriptional level. RNA helicases of the DEAD-box family play an important part in this process. In particular, CshA, which is part of the degradosome, is required for the rapid turnover of certain mRNAs and its deletion results in cold-sensitivity. To understand the molecular basis of this phenotype, we conducted a large genetic screen isolating 82 independent suppressors of cold growth. Full genome sequencing revealed the fatty acid synthesis pathway affected in many suppressor strains. Consistent with that result, sublethal doses of triclosan, a FASII inhibitor, can partially restore growth of a cshA mutant in the cold. Overexpression of the genes involved in branch-chained fatty acid synthesis was also able to suppress the cold-sensitivity. Using gas chromatography analysis of fatty acids, we observed an imbalance of straight and branch-chained fatty acids in the cshA mutant, compared to the wild-type. This imbalance is compensated in the suppressor strains. Thus, we reveal for the first time that the cold sensitive growth phenotype of a DEAD-box mutant can be explained, at least partially, by an improper membrane composition. The defect correlates with an accumulation of the pyruvate dehydrogenase complex mRNA, which is inefficiently degraded in absence of CshA. We propose that the resulting accumulation of acetyl- CoA fuels straight-chained fatty acid production at the expense of the branched ones. Strikingly, addition of acetate into the medium mimics the cshA deletion phenotype, resulting in cold sensitivity suppressed by the mutations found in our genetic screen or by sublethal doses of triclosan.
|
| |
|
| Overall design |
S. aureus is cold sensitive when deleting cshA gene. We produced 82 DcshA cold-suppressing strains and send them for whole genome sequencing in order to identify mutations involved in the supressing process. However, to limit library preparation cost, DNA samples were pooled into 9 groups of up to 12 DNA samples without barcoding.
|
| |
|
| Contributor(s) |
Khemici V, Prados J, Petrignani B, Di Nolfi B, Berge E, Manzano C, Giraud C, Linder P |
| Citation(s) |
32730248 |
| |
| Submission date |
Jun 19, 2019 |
| Last update date |
Aug 10, 2020 |
| Contact name |
Julien Prados |
| E-mail(s) |
julien.prados@unige.ch
|
| Phone |
+41 22 37 95 396
|
| Organization name |
University of Geneva
|
| Department |
SCMU
|
| Lab |
Bioinformatics Support Platform
|
| Street address |
Rue Michel Servet 1
|
| City |
Geneva |
| ZIP/Postal code |
1211 |
| Country |
Switzerland |
| |
|
| Platforms (1) |
| GPL19006 |
Illumina HiSeq 2500 (Staphylococcus aureus) |
|
| Samples (9)
|
| GSM3898309 |
GHU-55: Pool1 of 10 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898310 |
GHU-56: Pool2 of 10 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898311 |
GHU-57: Pool3 of 6 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898312 |
GHU-58: Pool4 of 8 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898313 |
GHU-59: Pool5 of 8 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898314 |
GHU-60: Pool6 of 7 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898315 |
GHU-61: Pool7 of 10 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898316 |
GHU-62: Pool8 of 12 genomic DNA sample from dcshA cold-suppressor strains |
| GSM3898317 |
GHU-63: Pool9 of 11 genomic DNA sample from dcshA cold-suppressor strains |
|
| Relations |
| BioProject |
PRJNA549691 |
| SRA |
SRP201896 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE133013_S_aureus_DcshA_suppressors.tsv.gz |
11.9 Kb |
(ftp)(http) |
TSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
|
|
|
|
 |
Less...
More...