Project description:We report a method to use BioID proximity labeling to map the sub-organization of translating mRNAs on the endoplasmic reticulum. This method first isolates membrane bound polysomes using a Sephacryl 400 resin-based enrichment followed by isolation of biotin labeled polysomes by streptavidin beads and high-throughput sequencing of the bound polysomes. We find that, for the two pools of polysomes labeled by either LRRC59 or Sec61beta BioID reporters, there is an interesting divergence in specific mRNAs that are entering into the secretory pathway such that secreted proteins are much more enriched towards the LRRC59 reporter with Sec61beta serving a more varied role in mRNA translation.
Project description:In this study, DYRK2 was found to be an important regulator of Hedgehog signaling. To identify DYRK2 interacting proteins, we performed interactome using BioID proximity labeling.
NIH3T3 cells containing Dox-inducible DYRK2-BioID2-HA construct were cultured in the media with or without Doxycycline for DYRK2-BioID2-HA expression, followed by proximity labeling. The lysates from Doxycycline induction (DYRK2) and without induction (Mock) were subjected to streptavidin pull-down and the proteins were identified by LC-MS/MS.
Project description:To gain insights into the interactome of wild-type (WT) and S102P mutant GATAD1, we utilized the BioID method, which enables the study of protein-protein interactions. Specifically, we performed BioID proximity labeling experiments in stable Flp-In cells expressing different GATAD1 variants fused to BirA*_FLAG. These variants included BirA*_FLAG_GATAD1-WT, BirA*_FLAG_GATAD1-S102P, BirA*_FLAG_GATAD1-S102D, and BirA*_FLAG_GATAD1-S102A. By employing this approach, we aimed to characterize the protein interactors associated with these GATAD1 variants and gain insights into the functional consequences of the S102P mutation.
Project description:Investigation of interactors for the human transcription factor PAX8 using BioID-based proximity labeling, as described in Galli et al.
Project description:To identify human DTX2 proximal proteome, the BioID-DTX2 fusion protein was expressed in DTX2 KO HeLa cells. The biotinylated proteins were pulled down and subjected to mass spectrometry analysis.
Project description:This study used the BioID proximity assay to establish the BACE1 interactome in healthy neuronal cells and identified interactions involved in BACE1 trafficking, post-translational modification and substrates.
Project description:This proof-of-principle experiment was designed to demonstrate the feasibility of proximity labeling for RNAM-bM-^@M-^Sprotein interactions IPL-seq on 293T-Rex expressing MSA-SNRPN70 (sample) or NFH-SNRPN70 (control)