Staufen protein is an RNA-binding protein that forms ribonucleoprotein complexes with the RNA, e.g., stress granules which help survive stress conditions. Changes of RNA-binding proteins can cause a number of neurodegenerative diseases characterised by neuronal degeneration. In our study, we used a proximity labelling method to label proteins that interact or are in the vicinity of the Staufen protein. Two biotin ligases were used: BirA* and TurboID. After confirming that the fusion protein was correctly expressed and that the biotinylation of the proteins was successful, we sent the isolated proteins for analysis by mass spectrometry. We concluded that most of the proteins we identified are involved in RNA binding or ribosome structure, and few proteins bind to DNA. We did not detect any major difference between the proteins surrounding Staufen 1 and Staufen 2 proteins. Using different biotin-ligases gave us insight into the temporal dynamics of the interactions. We showed that the Staufen protein is located near the the ribosome and the RNA molecule.
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