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Swiprosin1 controls cargo-specific entry to clathrin and dynamin-independent endocytic pathway

ABSTRACT: The clathrin and dynamin-independent endocytic pathway (CLIC-GEEC) is the major route of internalization of a large fraction of glycosylphosphatidylinositol-anchored proteins (GPI-APs) and other cell surface receptors. Unlike the clathrin-dependent endocytic pathway, where adaptors recruited to the endocytic pit provide the clues to recruit specific cargo, the CLIC-GEEC pathway is considered to take up portions of the membrane without any specificity. Here, we identified actin-binding protein swiprosin1 as the first cargo adaptor for the CLIC-GEEC pathway. We found that swiprosin1 interacts with active Rab21, a known regulator of integrin endocytosis, and couples Rab21 and the integrin cargo to the CLIC-GEEC endocytic machinery. Our work also identified the previously unknown pathway by which Rab21 mediates integrin endocytosis and linked the CLIC-GEEC pathway to integrin-dependent cancer cell invasion.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Henrik Zauber

LAB HEAD: Matthias Selbach

PROVIDER: PXD016478 | Pride | 2021-08-18

REPOSITORIES: Pride

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			Action	DRS
	MaxQuant_Output.zip	Other
	Statler_20150223_CHS_Veltel_A-H.raw	Raw
	Statler_20150223_CHS_Veltel_A-L.raw	Raw
	Statler_20150223_CHS_Veltel_B-H.raw	Raw
	Statler_20150223_CHS_Veltel_B-L.raw	Raw

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Project description:Graves’ ophthalmopathy (GO) is an extrathyroidal complication of Graves’ hyperthyroidism. Orbital fibroblast, an important cell in the pathogenesis of GO, is responsible for GO characteristics during active and inactive stages; however, information on the extensive profiles and the mechanistic regulations behind orbital fibroblast phenotypes are currently limited. The aim of this study was to compare the proteome profile of orbital fibroblasts isolated from inactive GO, active GO and healthy control. The proteome profile was further integrated with global DNA methylation data to identify this epigenetics regulation involved in the pathogenesis of GO. Methods Orbital fibroblasts culture isolated from five inactive GO, four active GO and five controls were cultured for total protein and DNA extraction. The labelled and fractionated proteins were analyzed with a liquid chromatography tandem-mass spectrometer (LC-MS/MS). On the other hand, the bisulphite-treated DNA was measured with the Illumina Infinium Human Methylation 450K beadchip. Proteome and methylation data were validated by real-time quantitative (RQ)-PCR. Network, pathway and functional analysis were performed by Ingenuity (Qiagen). Results Orbital fibroblasts from active GO displayed overexpression of proteins typically involved with inflammation, cellular proliferation, hyaluronan synthesis and adipogenesis, while several proteins linked to extracellular matrix (ECM) biology and fibrotic disease were overexpressed in orbital fibroblasts from inactive GO. The DNA methylation patterns were similar in healthy control and inactive GO orbital fibroblasts which were distinct from active GO orbital fibroblasts. In orbital fibroblasts from active GO hypermethylated genes link to inflammation while those genes hypomethylated are involved in adipogenesis and autoimmune-related genes. For the network analysis, overlapped between hypermethylated and hypomethylated genes were observed including NF-B, ERK1/2, Alp, RNA polymerase II, Akt and IFNα. In addition, NF-κB, Akt and IFNα were also identified in networks from the differentially expressed proteins. In general, poor correlation between protein expression, DNA methylation and mRNA expression in our study was observed.

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Swiprosin1 controls cargo-specific entry to clathrin and dynamin-independent endocytic pathway

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