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| Author(s): |
Alexandre Magno Vicente
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2017-02-17 |
| Examining board members: |
Marilis do Valle Marques;
Cristina Elisa Alvarez Martinez;
Carla Columbano de Oliveira
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| Advisor: | Marilis do Valle Marques |
| Abstract | |
Here, we constructed a strain in which the RhIE protein was fusioned to an epitope that allowed the investigation of the protein profile after cold-shock, at stationary phase, during cell cycle of Caulobacter crescentus, under different environmental stresses, and, finally, after chloramphenicol addition to study protein stability. The results showed an increase in protein concentration after cold shock stress. When exposed to different stresses RhlE was slightly induced in the presence of NaCl and Sucrose; but its abundance remained constant at stationary phase and during C. crescentus cell cycle. Lastly, the protein stability varies depending on temperature - increasing at low temperature. Gene expression analysis was performed using beta-galactosidase assays. We showed that the presence of the 5UTR is important for the induction of rhlE, and that RhlE is posttranscriptionally regulated. A global transcriptome analysis was performed using RNAseq after cold shock stress of the wild type strain and null mutant for rhlE, and several genes involved in a wide range of biological process were identified. Finally, High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation revealed interactions of RhlE with 51 mRNAs. (AU) | |
| FAPESP's process: | 14/13552-4 - Study of the DEAD-box RNA helicase encoded by the gene CC0835 in Caulobacter crescentus |
| Grantee: | Alexandre Magno Vicente |
| Support Opportunities: | Scholarships in Brazil - Master |