Identification of BOC proximity proteins by BioID
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Q Exactive HF
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: 
Frederic Charron
PROVIDER: MSV000091742 | MassIVE | Wed Apr 19 07:18:00 BST 2023
SECONDARY ACCESSION(S): PXD041651
REPOSITORIES: MassIVE
Similar Datasets
Project description:In the developing spinal cord, Sonic hedgehog (Shh) attracts commissural axons toward the floor plate. How Shh regulates the cytoskeletal remodeling that underlies growth cone turning is unknown. We found that Shh-mediated growth cone turning requires the activity of Docks, which are unconventional GEFs. Knockdown of Dock3 and 4, or their binding partner ELMO1 and 2, abolished commissural axon attraction by Shh in vitro. Dock3/4 and ELMO1/2 were also required for correct commissural axon guidance in vivo. Polarized Dock activity was sufficient to induce axon turning, indicating that Docks are instructive for axon guidance. Mechanistically, we show that Dock and ELMO interact with Boc, the Shh receptor, and that this interaction is reduced upon Shh stimulation. Furthermore, Shh stimulation translocates ELMO to the growth cone periphery and activates Rac1. This identifies Dock/ELMO as an effector complex of non-canonical Shh signaling and demonstrates the instructive role of GEFs in axon guidance.
2024-05-31 | GSE268644 | GEO
Project description:Proximity dependent biotin identification (BioID) has emerged as a powerful tool for studies of proteome architecture, including insoluble or membrane-associated proteins. The technique has been well established in mammalian cells but has yet to be applied to whole plant systems. Here we demonstrate the application of BioID on leaf tissues of the model plant Arabidopsis thaliana, thereby expanding the versatility of this important technique and providing a powerful proteomics tool for plant biologists.
| S-EPMC6004002 | biostudies-literature
Project description:Mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) is Ser/Thr protein kinase highly expressed in medulloblastoma (MB) patient samples which promotes cell migration and invasiveness downstream of growth factor receptors. Although MAP4K4 implication in the control of the membrane protrusions at the leading edge of cells during migration and invasion in MB is established, its downstream effectors governing and mediating this process in MB have not been identified. In this study we explored the proteomic neighborhood of MAP4K4 in MB cells and to identify candidate potentially affecting its activity and pro-migratory functions. Towards this aim, we expressed BioID fused versions of MAP4K4 (N- and C-terminal) in DAOY cells under presence of biotin. In the control setup, we expressed the BioID enzyme alone. After Streptavidin purification of biotinylated proteins, we performed mass spectrometry and quantified relative abundances compared to control conditions.
2022-08-04 | PXD031870 | Pride
Project description:Mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) is Ser/Thr protein kinase highly expressed in medulloblastoma (MB) patient samples which promotes cell migration and invasiveness downstream of growth factor receptors. Although MAP4K4 implication in the control of the membrane protrusions at the leading edge of cells during migration and invasion in MB is established, its downstream effectors governing and mediating this process in MB have not been identified. In this study we explored the proteomic neighborhood of MAP4K4 in MB cells and to identify candidate potentially affecting its activity and pro-migratory functions. Towards this aim, we expressed BioID fused versions of MAP4K4(N- and C-terminal) in HEK-293T cells under presence of biotin. In the control setup, we expressed the BioID enzyme alone. After Streptavidin purification of biotinylated proteins, we performed mass spectrometry and quantified relative abundances compared to control conditions.
2022-08-04 | PXD031863 | Pride
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.
2022-10-11 | PXD023795 |
Project description:Reported here are the transcriptomic changes resulting from learning, in the brain of an anuran amphibian. RNA sequencing was utilized to investigate temporal gene expression patterns resulting from conditioned learning in the fire-bellied toad, Bombina orientalis. As a basal tetrapod lineage, B. orientalis represents a robust candidate for investigating the evolution of molecular mechanisms underlying learning in vertebrates. Differential expression following learning reveled activity in molecular pathways related to immediate early gene expression, cytoskeletal modification, axon guidance activity, and apoptotic processes. Learning in the fire-bellied toad is characterized by early IEG activity coinciding with transcriptional activity and structural modification, followed by axon guidance and adhesion activity, and late neural pruning.
2020-01-01 | GSE135054 | GEO
Project description:The BioID method uses a promiscuous biotin ligase to detect protein-protein associations as well as proximate proteins in living cells. Here we report improvements to the BioID method centered on BioID2, a substantially smaller promiscuous biotin ligase. BioID2 enables more-selective targeting of fusion proteins, requires less biotin supplementation, and exhibits enhanced labeling of proximate proteins. Thus BioID2 improves the efficiency of screening for protein-protein associations. We also demonstrate that the biotinylation range of BioID2 can be considerably modulated using flexible linkers, thus enabling application-specific adjustment of the biotin-labeling radius.
| S-EPMC4831873 | biostudies-literature
Project description:We have discovered subsets of axon guidance molecules and transcription factors that are enriched in specific subsets of olfactory sensory neurons. We have demonstrated guidance activity for three of the candidate axon guidance genes we identified, suggesting that this approach is an efficient method for characterizing guidance systems relevant to olfactory axon targeting.
2018-02-19 | GSE103692 | GEO
Project description:During the development of the Drosophila central nervous system the process of midline crossing is orchestrated by a number of guidance receptors and ligands. Many key axon guidance molecules have been identified in both invertebrates and vertebrates, but the transcriptional regulation of growth cone guidance remains largely unknown. One open question is whether transcriptional regulation plays a role in midline crossing, or if local translation can account for the necessary fine tuning of protein levels. To investigate this issue, we conducted a genome wide analysis of transcription in Drosophila embryos using wild type and a number of well-characterized Drosophila guidance mutants and transgenics. We also analyzed a publicly available microarray time course of Drosophila embryonic development with an axon guidance focus. Using hopach, a novel clustering method which is well suited to microarray data analysis, we identified groups of genes with similar expression patterns across guidance mutants and transgenics. We then systematically characterized the resulting clusters with respect to their relevance to axon guidance using two complementary controlled vocabularies: the Gene Ontology (GO) and anatomical annotations of the Atlas of Pattern of Gene Expression (APoGE) in situ hybridization database. The analysis indicates that regulation of gene expression does play a role in the process of axon guidance in Drosophila. We also find a strong link between axon guidance and hemocyte migration, a result that agrees with mounting evidence that axon guidance molecules are co-opted in vertebrate vascularization. Cell cyclin activity in the context of axon guidance is also suggested from our array data. RNA and protein patterns of cell cyclin in axon guidance mutants and transgenics support this possible link. This study provides important insights into the regulation of axon guidance in vivo and suggests that transcription does play a role in control of axon guidance. Keywords: Mutant Analysis
2006-12-14 | GSE6515 | GEO
Project description:During the development of the Drosophila central nervous system the process of midline crossing is orchestrated by a number of guidance receptors and ligands. Many key axon guidance molecules have been identified in both invertebrates and vertebrates, but the transcriptional regulation of growth cone guidance remains largely unknown. One open question is whether transcriptional regulation plays a role in midline crossing, or if local translation can account for the necessary fine tuning of protein levels. To investigate this issue, we conducted a genome wide analysis of transcription in Drosophila embryos using wild type and a number of well-characterized Drosophila guidance mutants and transgenics. We also analyzed a publicly available microarray time course of Drosophila embryonic development with an axon guidance focus. Using hopach, a novel clustering method which is well suited to microarray data analysis, we identified groups of genes with similar expression patterns across guidance mutants and transgenics. We then systematically characterized the resulting clusters with respect to their relevance to axon guidance using two complementary controlled vocabularies: the Gene Ontology (GO) and anatomical annotations of the Atlas of Pattern of Gene Expression (APoGE) in situ hybridization database. The analysis indicates that regulation of gene expression does play a role in the process of axon guidance in Drosophila. We also find a strong link between axon guidance and hemocyte migration, a result that agrees with mounting evidence that axon guidance molecules are co-opted in vertebrate vascularization. Cell cyclin activity in the context of axon guidance is also suggested from our array data. RNA and protein patterns of cell cyclin in axon guidance mutants and transgenics support this possible link. This study provides important insights into the regulation of axon guidance in vivo and suggests that transcription does play a role in control of axon guidance. Experiment Overall Design: Several mutants and transgenics were analyzed, totalizing 17 distinct conditions. Individual descriptions are included in each microarray. For each condition there are 3 or 4 replicates. The file's name indicates the replicates, e.g., comm.a reflects the replicate a of mutant commissureless. The experiment design covers a range of mutants and transgenics of key axon guidance mutans, in different dosages. The protocol was the standard Affymetrix protocol.
2008-06-15 | E-GEOD-6515 | biostudies-arrayexpress