Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 23 for “"Proximity labelling"”.
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INTERACTOMICS FOR RICE FLOWERING: A PROXIMITY LABELLING APPROACH
… nel nostro laboratorio, abbiamo utilizzato il proximity labelling (PL) per esplorare le interazioni di Hd1 e FTL1 con altre proteine. Il PL si basa sulla fusione della TurboID (TbID), una biotin-ligasi ottimizzata, alla proteina di interesse con lo scopo di marcarne gli interattori in vivo. …
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Exploring Polarized Membrane Protein Distribution in Drosophila Follicle Cells using Proximity Labelling
… model, this thesis applies peroxidase-mediated proximity labelling and mass spectrometry to systematically characterize the polarized membrane proteome. I first demonstrate that peroxidase-catalyzed labelling with biotin-phenol derivatives is a diffusion-controlled process: excess phenoxy …
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Investigation towards the role of LmxOSM3.1 phosphorylation and the identification of its interaction partners using an improved method of proximity labelling
… they coordinate flagellum length.;In 2C-mTID, a proximity-based assay to define potential protein-protein interactions (PPIs), miniTurbo (mTID) and LmxOSM3.1 (PoI) are fused to FKBP and FRB (of the FRB-FKBPmammalian oligomerisation system), respectively. A cell line expressing the CRISPR-Cas9 …
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Identification and characterisation of novel interactors of surfactant protein C
… the I73T variant, I undertook SFTPC-WT and -I73T proximity labelling using TurboID and identified labelled proteins by mass spectrometry. I discovered >100 proteins preferentially labelled by SFTPC-WT and therefore likely to be required for normal SFTPC trafficking. I undertook validation of …
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Elucidation and characterisation of checkpoint kinase interactions with the replisome
… identification technique called biotin proximity labelling to elucidate novel interactors of the budding yeast checkpoint kinase Rad53 with the replisome. Using this technique, Pol1, a polymerase involved in DNA replication, was found to be a putative Rad53 interactor. Pol1 interacts …
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Characterizing the Mechanisms Regulating Enhancer Commissioning by MyoD in Muscle Cells
… is regulated in muscle cells, we first utilize proximity labelling coupled with mass spectrometry to identify potential coactivators of MyoD. Our results reveal a link between MyoD and the Mediator complex, a key component of enhancer-promoter looping. Next, using phenotypic assays and …
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Investigating how mitophagy is regulated by the metabolic context
… a proteomic approach using APEX2 biotin proximity labelling to detect changes in mitophagy signalling from the mitochondria surface. I exploited the in vitro and in vivo validated mito-QC reporter to target APEX2 on the mitochondrial surface using a conditionally stable anti-GFP nanobody. …
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Mapping the cell cycle-dependent centrosomal interactome
… anchored on the centrosome, an endogenous proximity labelling system called miniTurbo was fused with centrosomal proteins Centrin 2, PCNT and Cep192 to detect the centrosome interactome. The cell cycle-dependent interactome of the centrosome was mapped by combining proximity biotinylation …
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Regulatory mechanisms of MYO6 motor domain activity and tail interactions
… sites. Lastly, recent advances in proximity labelling techniques allow me to determine dynamic MYO6 interactions *in vivo*, at steady state conditions and after the induction of autophagy. Overall, this study provides novel insights into the dynamic regulation of MYO6 function and …
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Identification and functional characterisation of synaptic AMPA receptor-interacting proteins
… proteins. Aided by the development of a novel proximity-labelling proteomics technique, synaptic proteins enriched for the AMPAR NTD were determined. This permitted identification of transient NTD interactors, previously unresolved using classical affinity-purifications. Furthermore, this …
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The role of FOXK transcription factors in Wnt signal transduction
… I employed a proteomics approach based on proximity labelling (BioID) using TLE3 and Pygopus as bait proteins. This revealed little changes of the core enhanceosome subunits between active or inactive signalling but indicated the Forkheadbox transcription factors FOXK1/2 as potential …
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Investigating the molecular environments and interactions of pathological TDP-43 filaments
… I developed a time-resolved, antibody-targeted proximity labelling approach to map the molecular environments and potential interactions of pathological TDP-43 in these models at different stages of propagation in an unbiased manner. During the uptake of exogenous filaments, I observed an …
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Study of the intracellular trafficking of the pre-T-cell receptor and its role in T-cell lineage commitment
… down the γδ-lineage. Finally, I employ a biotin proximity-labelling assay to screen for binding partners of the pre-TCR that might mediate its internalisation. Using this method, I identify TMEM131, a novel protein with an unknown function and characterise some of its biochemical and cell …
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Molecular Characterisation and Therapeutic Targeting of NPM1-mutant Acute Myeloid Leukaemia
… we designed and initiated the development of a proximity-labelling platform in NPM1-mutant AML cells. However, a series of functional assays revealed that lentivirally-delivered NPM1c fusion proteins do not mimic the behaviour of the native protein; results with potential implications for future …
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A New Approach To Identifying Effectors and Regulators of Small GTPases
… an adaptation of the recently developed in vivo proximity labelling technique BioID was used in this study. Here, BioID was redirected to the ectopic location of the surface of mitochondria (hereafter called MitoID) with the aim of restricting the non-specific background. Applying this method to …
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The Role of Autotaxin in Cancer
… protein-protein interactions. An HRP-based proximity labelling method was optimised for investigating the extracellular protein interactors of ATX. Several potential interactors are identified, including fatty acid binding protein 5 (FABP5). Modelling with AlphaFold-Multimer indicates FABP5 …
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Characterisation of LITAF, a protein associated with Charcot-Marie-Tooth disease type 1C
… traditional GST pull-down assay and the in-situ proximity labelling assay, BioID. A number of novel potential binding partners were identified in both assays. Among the list of potential binding partners, BAG3 was captured in both pull-down assays and was chosen for further studies. The …
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LOCALIZED WNT3A-SOURCES INSTRUCT MITOTIC ORIENTATION: DISSECTING THE WNT3A-TRANSDUCTION MECHANISM COUPLED WITH NUMA MACHINERY
… for Wnt3a-dependent spindle orientation. Proximity labelling assay revealed that during mitosis the Wnt3a co-receptor LRP6 is in proximity of NuMA and Gai subunits. Moreover, performing immunoprecipitation assays we demonstrate that Wnt3a stimulation triggers the formation of …
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Functional Characterisation of Protein Degradation during Human Cytomegalovirus Infection
… interactors of neurabin-II were identified by proximity-labelling mass spectrometry, however all candidates appeared to be dispensable for neurabin-II downregulation and could not elucidate the viral mechanism of degradation. Chapter four utilised the neddylation inhibitor MLN4924 which impairs …
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AN INVESTIGATION OF THE MECHANISMS OF GROUCHO MEDIATED TRANSCRIPTIONAL REPRESSION
… yeast 2-hybrid screen, and in a TurboID-based proximity labelling assay in Drosophila Kc167 cells. However, these approaches resulted in unfruitful finding novel interacting partners. Taking together all the evidence from this work I proposed a model of Gro-mediated repression, in which Gro …
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