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| Status |
Public on Nov 03, 2017 |
| Title |
Cold adaptation of tRNA nucleotidyltransferases: a tradeoff in activity, stability and fidelity |
| Organisms |
Geobacillus stearothermophilus; Bacillus subtilis subsp. subtilis str. 168; Exiguobacterium artemiae; Planococcus halocryophilus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
CCA-adding enzymes are highly specific RNA polymerases that synthesize and maintain the sequence CCA at the tRNA 3‘-end. Here, we investigated the impact of cold adaptation on the reactivity and specificity of CCA-adding enzymes from psychrophilic bacteria. A comparative study of the corresponding enzymes from closely related psychro-, meso-, and thermophilic Bacillales indicates that the cold-adapted enzymes show a considerable error rate during CCA synthesis, resulting in additional incorporations of C and A residues. It seems that the activity of psychrophilic CCA-adding enzymes is not only achieved at the expense of structural stability, reaction velocity and substrate affinity, but also results in a reduced polymerization fidelity.
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| Overall design |
in vivo tRNA from four Bacillales species was isolated from exponential and stationary growth phase each in triplicates resulting in a total of 24 samples. Sequencing libraries were created for small RNA species and sequenced on 3 runs of a MiSeq using 8 indices. Reads were analyzed regarding 3'-ends of tRNA and results summed up as a mean for the 8 conditions investigated. Results from B. subtilis were regarded as reference sample for a mesophilic Bacillales species. Differences towards psychophilic Planococcus halocryophilus and Exiguobacterium sibiricum and thermophilic Geobacills stearothermophilus were investigated.
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| Contributor(s) |
Ernst FG, Juehling F, Moerl M |
| Citation(s) |
29099323 |
| |
| Submission date |
May 12, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Felix Georg Maria Ernst |
| Organization name |
University of Leipzig
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| Department |
Faculty of Bioscience
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| Lab |
Molcular Biology
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| Street address |
Bruederstrasse 34
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| City |
Leipzig |
| ZIP/Postal code |
04103 |
| Country |
Germany |
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| Platforms (4)
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| GPL23470 |
Illumina MiSeq (Planococcus halocryophilus) |
| GPL23471 |
Illumina MiSeq (Exiguobacterium sibiricum) |
| GPL23472 |
Illumina MiSeq (Geobacillus stearothermophilus) |
| GPL23473 |
Illumina MiSeq (Bacillus subtilis subsp. subtilis str. 168) |
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| Samples (23)
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| Relations |
| BioProject |
PRJNA386487 |
| SRA |
SRP106999 |
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