Browse
Submit Data
Databases
API
Help

Dataset Information

242 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

The MYO6 interactome using BioID

ABSTRACT: The intracellular function of myosin motors requires a number of adaptor molecules, which control cargo attachment, but also fine-tune motor activity in time and space. These motor-adaptor-cargo interactions are often weak, transient or highly regulated. To overcome these problems we use a proximity labelling-based proteomics strategy (BioID) to map the interactome of the unique minus end-directed actin motor MYO6. Our analysis identified several distinct MYO6-adaptor modules including two complexes containing RHO GEFs which we screened using further BioID baits (LRCH3, DOCK7, GIPC1 and ARHGEF12). These complexes emphasise the multifunctionality of MYO6 provides the first in vivo interactome of a myosin motor protein, highlighting the power of this approach in uncovering dynamic and functionally diverse myosin motor complexes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Retinal Pigment Epithelial Cell

DISEASE(S): Disease Free

SUBMITTER: Tom O'Loughlin

LAB HEAD: Dr. Folma Buss

PROVIDER: PXD008686 | Pride | 2018-02-27

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	BirA_190517_070617.raw	Raw
	BirA_27336.raw	Raw
	BirA_27337.raw	Raw
	BirA_27338.raw	Raw
	BirA_280517_070617.raw	Raw

Items per page:

1 - 5 of 39

Publications

The MYO6 interactome reveals adaptor complexes coordinating early endosome and cytoskeletal dynamics.

O'Loughlin Thomas T Masters Thomas A TA Buss Folma F

EMBO reports 20180221 4

The intracellular functions of myosin motors requires a number of adaptor molecules, which control cargo attachment, but also fine-tune motor activity in time and space. These motor-adaptor-cargo interactions are often weak, transient or highly regulated. To overcome these problems, we use a proximity labelling-based proteomics strategy to map the interactome of the unique minus end-directed actin motor MYO6. Detailed biochemical and functional analysis identified several distinct MYO6-adaptor m ...[more]

PMID: 29467281

Similar Datasets

UNC-45 assisted myosin folding depends on a conserved FX3HY motif implicated in Freeman Sheldon Syndrome

Project description:Myosin motors are critical for diverse motility functions ranging from cytokinesis and endocytosis to muscle contraction. The UNC-45 chaperone controls myosin function mediating the folding, assembly, and degradation of the muscle protein. Here, we analyze the molecular mechanism of UNC-45 as a hub in myosin quality control. We show that UNC-45 forms discrete complexes with folded and unfolded myosin, forwarding them to downstream chaperones and E3 ligases. Structural analysis of a minimal chaperone:substrate complex reveals that UNC-45 binds to a conserved FX3HY motif in the myosin motor domain. Disrupting the observed interface by mutagenesis prevents myosin maturation leading to protein aggregation in vivo. We also show that a mutation in the FX3HY motif linked to the Freeman Sheldon Syndrome impairs UNC-45 assisted folding, reducing the level of functional myosin. These findings demonstrate that a faulty myosin quality control is a critical yet unexplored cause of human myopathies.

2024-07-25 | PXD040428 | Pride

RNA-BioID of lncRNA HOTAIR in Flp-In T-REx cells

Project description:Accumulating evidence highlights the role of long non-coding RNAs (lncRNA) in cellular homeostasis, and their dysregulation in disease settings. Most lncRNAs function by interacting with proteins or protein complexes. While several orthogonal methods have been developed to identify these proteins, each method has its inherent strengths and limitations. Here, we combine two RNA-centric methods ChIRP-MS and RNA-BioID to obtain a comprehensive list of proteins that interact with the well-known lncRNA HOTAIR. Overexpression of HOTAIR has been associated with a metastasis-promoting phenotype in various cancers. Although HOTAIR is known to bind with PRC2 and LSD1 protein complexes, an unbiased and comprehensive method to map its interactome has not yet been performed. Both ChIRP-MS and RNA-BioID data sets show an association of HOTAIR with mitoribosomes, suggesting HOTAIR has functions independent of its (post-)transcriptional mode-of-action.

2022-02-15 | PXD029058 | Pride

Active Promoters give rise to false positive 'Phantom Peaks' in ChIP-Seq experiments

Project description:We applied ChIP-seq to map the chromosomal binding sites for two nucleosome remodeling complexes containing the ATPase ISWI, ACF and RSF, in Drosophila embryos. Employing a panel of polyclonal and monoclonal antibodies directed against their signature subunits, ACF1 and RSF1, robust profiles were obtained indicating that both remodelers co-occupied a large set of active promoters. For further validation we repeated the mapping using chromatin of mutant embryos that do not express ACF1 or RSF1. Surprisingly, the ChIP-seq profiles were unchanged, suggesting that they were not due to specific immunoprecipitation. Conservative analysis lists about 3000 chromosomal loci, mostly active promoters that are prone to non-specific enrichment in ChIP and give rise to ‘Phantom Peaks’. These peaks are not obtained with pre-immune serum and are not prominent in input chromatin. Examination of various ACF1 and RSF1 antibodies in Drosophila melanogaster embryos which are wildtype or mutant for the antibody targets.

2015-07-10 | E-GEOD-67323 | biostudies-arrayexpress

Chromatin immunoprecipitation of mouse polycomb group (PcG) proteins in embryonic stem cells

Project description:Polycomb group (PcG) proteins are highly conserved from flies to mammals and many of these factors have essential roles in early embryonic development. PcG proteins comprise two multimeric complexes, the Polycomb Repressive complexes 1 and 2 (PRC1 and 2), which have been shown to repress transcription through epigenetic modification of chromatin structure. To gain insight into the role of Polycomb in early development, we have identified PRC1 and PRC2 target genes in mouse embryonic stem (ES) cells using genome-scale location analysis. We found that PRC2 occupies many genes that encode key regulators of development including those encoding transcription factors and components of signaling pathways. These genes are repressed and contain nucleosomes methylated at lysine 27 on histone H3. The majority of PRC2 bound and methylated target genes are co-occupied by PRC1 indicating that these complexes function at a similar set of genes in ES cells. Lack of PRC1 or PRC2 subunits in ES cells results in derepression of target genes and loss of pluripotency. These results provide insight into how PcG proteins contribute to the maintenance of stem cell identity.

2006-05-09 | E-WMIT-12 | biostudies-arrayexpress

R2TP_BioID-MS - R2TP/Prefoldin-like component RUVBL1/RUVBL2 directly interacts with ZNHIT2 to regulate assembly of U5 small nuclear ribonucleoprotein

Project description:The R2TP/Prefoldin-like (R2TP/PFDL) complex is a cochaperone complex involved in the assembly of a number of critical protein complexes. Here we use proximity-dependent biotinylation (BioID) coupled to mass spectrometry to confirm interactions of ZNHIT2, a cofactor of R2TP/PFDN, identified by TAP-MS.

2018-10-24 | PXD006200 | Pride

Caldendrin regulates spine apparatus localization through myosin V-dependent ER-stabilization in dendritic spines

Project description:Excitatory synapses of principal hippocampal neurons are frequently located on dendritic spines. The dynamic strengthening or weakening of individual inputs results in a great structural and molecular diversity of dendritic spines. Active spines with large Ca2+-transients are frequently invaded by a single protrusion from the endoplasmic reticulum (ER), which is dynamically transported into spines by the actin-based motor myosin V. An increase in synaptic strength often correlates with stable anchoring of the ER, followed by the formation of the spine apparatus organelle. Here we characterize the newly identified interaction of myosin V with the Ca2+-sensor caldendrin, a brain-specific homolog of the well-known myosin V interactor calmodulin. While calmodulin is an essential activator of myosin V motor function, we found that caldendrin acts as an inhibitor of processive myosin V movement. We propose that caldendrin transforms myosin into a stationary F actin tether, and show that in mouse and rat hippocampal neurons, caldendrin regulates spine apparatus localization to a subset of dendritic spines through a myosin V-dependent pathway.

2022-01-27 | PXD029427 | Pride

Mapping myddosome components in human and murine macrophages.

Project description:Toll-like Receptors (TLRs) are key mediators of immune responses to infection. Upon microbial detection, these receptors activate inflammatory signal transduction pathways that involve IκB kinases, mitogen activated protein kinases, ubiquitin ligases and other adaptor proteins. Current models suggest these signaling proteins operate within functionally distinct multiprotein complexes, which are all activated by a receptor-proximal complex known as the myddosome. The mechanisms that connect the effector protein complexes in the TLR pathways are undefined. To define TLR pathway activities, we engineered macrophages to enable proteomic analysis of the endogenous myddosome constituent MyD88 and its interacting proteins. Proteomics demonstrated that myddosomes contain proteins that act at all stages and regulate all effector responses of the TLR signaling pathways.

2024-07-04 | PXD047416 | Pride

The OPTN interactome using BioID

Project description:We use a proximity labelling-based proteomics strategy (BioID) to map the interactome of OPTN. OPTN was tagged N and C-terminally with BirA* in RPE1 cells and then, after labelling with biotin, streptavidin pull downs were performed to identify OPTN-proximal proteins.

2020-06-16 | PXD019386 | Pride

Epistasis studies reveal redundancy among calcium-dependent protein kinases in motility and invasion of malaria parasites

Project description:In malaria parasites, evolution of parasitism has been linked to functional optimisation. Despite this optimisation, most members of a calcium-dependent protein kinase (CDPK) family show genetic redundancy during erythrocytic proliferation. To identify relationships between phospho-signalling pathways, we here screen 294 genetic interactions among protein kinases in Plasmodium berghei. This reveals a synthetic negative interaction between a hypomorphic allele of the protein kinase G (PKG) and CDPK4 to control erythrocyte invasion which is conserved in P. falciparum. CDPK4 becomes critical when PKG-dependent calcium signals are attenuated to phosphorylate proteins important for the stability of the inner membrane complex, which serves as an anchor for the acto-myosin motor required for motility and invasion. Finally, we show that multiple kinases functionally complement CDPK4 during erythrocytic proliferation and transmission to the mosquito vector. This study reveals how CDPKs are wired within a stage-transcending signalling network to control motility and host cell invasion in malaria parasites.

2018-09-17 | PXD011096 | Pride

A farnesyl-dependent structural role for CENP-E in expansion of the fibrous corona

Project description:Correct chromosome segregation during cell division depends on proper connections between spindle microtubules and kinetochores. During prometaphase, kinetochores are temporarily covered with a dense protein meshwork known as the fibrous corona. Formed by oligomerization of ROD/ZW10/ZWILCH-SPINDLY (RZZ-S) complexes, the fibrous corona promotes spindle assembly, chromosome orientation and spindle checkpoint signaling. The molecular requirements for formation of the fibrous corona are not fully understood. Here we show that the fibrous corona depends on the mitotic kinesin CENP-E, and that poorly expanded fibrous coronas after CENP-E depletion are functionally compromised. This previously unrecognized role for CENP-E does not require its motor activity but instead is driven by its C-terminal microtubule-binding region and farnesyl modification thereof. The ‘birth’ of a farnesylation motif in CENP-E orthologs during Obazoa evolution closely aligns with presence of the full RZZ-S module, and we show that in cells CENP-E interacts with RZZ-S complexes in a farnesyl-dependent manner. CENP-E is recruited to kinetochores following RZZ-S, and - while not required for RZZ-S oligomerization per se - promotes subsequent fibrous corona expansion. Our results show that proper spindle assembly has a non-motor contribution from the kinesin CENP-E through stabilization of the fibrous corona meshwork during its formation.

2023-10-09 | PXD040338 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data