Kinetics of rat CSD-C2 binding to H3.3 RNA
| Autor: | Saladino P, Gygax D, Spies P, Schiera G, Di Liegro I, Di Liegro C.M. |
|---|---|
| Přispěvatelé: | Saladino, P., Gygax, D., Spies, P., Schiera, G., Di Liegro, I., Di Liegro, C. |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Chemistry
0206 medical engineering Kinetics RNA 02 engineering and technology 021001 nanoscience & nanotechnology 020601 biomedical engineering Settore BIO/10 - Biochimica Automotive Engineering Biophysics RNA-protein interactions CSD-C2 Histone H3.3 RNA Biolayer interferometry Settore BIO/06 - Anatomia Comparata E Citologia 0210 nano-technology |
| Popis: | Cold-shock domain containing protein C2 (CSD-C2; also known as PIPPin) is an RNA-binding protein conserved in the evolution that interacts with the 3’-untranslated region (3’-UTR) of rat H1.0 and H3.3 histone messengers. Biolayer interferometry (BLI) is a technique that measures changes in an interference pattern generated from visible light, reflected from an optical layer, and a biolayer which contains molecules of interest. In this study, we used the BLI methodology in order to analyze and describe the binding properties of CSD-C2 and the mRNA encoding the rat brain histone protein H3.3. Recombinant CSD-C2 was incubated with in vitro transcribed, and biotinylated H3.3 RNA fragments bound to streptavidin-conjugated Octet optical biosensors. In order to define the RNA region involved in binding, we used RNA probes corresponding to different portions of H3.3 RNA 3’-UTR. In this study, we showed that CSD-C2 binds to the last 199 nucleotides of the H3.3 RNA 3’-UTR, and that the apparent affinity constant of the interaction is in the nanomolar range. In addition, this study confirmed that BLI can be a very efficient and reliable method for studying RNA-protein interactions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|