6533b82dfe1ef96bd1291f73
RESEARCH PRODUCT
Kinetics of rat CSD-C2 binding to H3.3 RNA
Saladino PGygax DSpies PSchiera GDi Liegro IDi Liegro C.m.subject
Chemistry0206 medical engineeringKineticsRNA02 engineering and technology021001 nanoscience & nanotechnology020601 biomedical engineeringSettore BIO/10 - BiochimicaAutomotive EngineeringBiophysicsRNA-protein interactions CSD-C2 Histone H3.3 RNA Biolayer interferometry.Settore BIO/06 - Anatomia Comparata E Citologia0210 nano-technologydescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2017-04-24 |