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| Status |
Public on Nov 25, 2019 |
| Title |
Pleiotropic effects of the rne∆MTS allele on RNA degradation |
| Platform organism |
Escherichia coli K-12 |
| Sample organism |
Escherichia coli str. K-12 substr. MG1655 |
| Experiment type |
Expression profiling by array
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| Summary |
Localization of RNase E to the inner membrane in Escherichia coli is well documented, but the functional consequences of this localization are largely unknown. Here we characterize the rne∆MTS strain, which expresses cytoplasmic RNase E (cRNase E). CsrB and CsrC regulatory RNAs are stabilized in the rne∆MTS strain resulting in leaky glycogen expression. There is a small but significant global slowdown in mRNA degradation with no bias considering function or localization of encoded proteins. RNase E is a stable protein, but cRNase E is unstable with a half-life equal to the doubling time of exponentially growing cells. cRNase E instability is compensated by increased synthesis. Co-purification experiments show that cRNase E associates with RhlB, enolase and PNPase to form a cytoplasmic RNA degradosome. Measurements in multiple turnover assays show that there is no difference in Km or kcat between cRNase E and RNase E. In contrast to the global slowdown of mRNA degradation, the inactivation of a ribosome-free lacZ transcript is faster in the rne∆MTS strain. We discuss how the association of RNase E with the inner cytoplasmic membrane is important for carbon storage regulation, degradation of polyribosomal mRNA, protection of ribosome-free transcripts from inactivation and stability of RNase E.
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| Overall design |
A microarray study was performed to estimate genome-wide mRNA half-lives in two E. coli K12 MG1655 strains, rne+ and rne(ΔMTS), cultured in M9+glucose (3g/l). In mid-exponential phase, sampling was before the addition of rifampicin (500 µg/l) for transcription arrest and over the time after rifampicin addition. Total RNA was then extracted from each sample. Each array measures the expression level of 4,254 genes from Escherichia coli MG1655 with eight 60-mer probes per gene in duplicates. Three independent kinetics were performed for each strain.
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| Contributor(s) |
Hadjeras L, Poljak L, Bouvier M, Morin-Ogier Q, Canal I, Cocaign-Bousquet M, Girbal L, Carpousis AJ |
| Citation(s) |
30903628 |
| |
| Submission date |
Aug 02, 2018 |
| Last update date |
Nov 25, 2019 |
| Contact name |
Laurence Girbal |
| E-mail(s) |
girbal@insa-toulouse.fr
|
| Phone |
33 5 61 55 97 24
|
| Organization name |
TBI
|
| Street address |
135 avenue de rangueil
|
| City |
Toulouse |
| ZIP/Postal code |
31077 |
| Country |
France |
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| Platforms (2) |
| GPL14649 |
NimbleGen E. coli K12 Gene Expression Array [071112_Ecoli_K12_EXP] |
| GPL25406 |
NimbleGen E. coli K12 Gene Expression Array [130108_Ecoli_K12_exp] |
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| Samples (24)
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| Relations |
| BioProject |
PRJNA484207 |
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