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Rap1a GTPase mVSM Knockout vs. Rescue miRNA Expression

ABSTRACT: miRNA profiling of mouse tail artery explanted microvascular smooth muscle (mVSM) cells comparing Rap1a -/- mVSM cells trasduced with vector backbone (control) with Rap1a -/- mVSM cells transduced with a Rap1 CA 'rescue' plasmid. The experiment will compare miRNA expression in Rap1A -/- mVSMs and 'rescued'with Rap1A CA.

ORGANISM(S): Mus musculus

PROVIDER: GSE24749 | GEO | 2011/04/13

SECONDARY ACCESSION(S): PRJNA142715

REPOSITORIES: GEO

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Rap1a GTPase mVSM Knockout vs. Rescue mRNA Expression

Project description:Transcriptional profiling of mouse tail artery explanted microvascular smooth muscle (mVSM) cells comparing Rap1a -/- mVSM cells transduced with vector backbone (control) with Rap1a -/- mVSM cells transduced with a Rap1 CA 'rescue' plasmid. The experiment will compare mRNA expression in Rap1A -/- mVSMs and 'rescued'with Rap1A CA.

2011-04-13 | GSE25702 | GEO

Rap1a GTPase mVSM Knockout vs. Rescue miRNA Expression

Project description:miRNA profiling of mouse tail artery explanted microvascular smooth muscle (mVSM) cells comparing Rap1a -/- mVSM cells trasduced with vector backbone (control) with Rap1a -/- mVSM cells transduced with a Rap1 CA 'rescue' plasmid. The experiment will compare miRNA expression in Rap1A -/- mVSMs and 'rescued'with Rap1A CA. Two-condition experiment, mVSM Rap1a -/- (vector control) vs. mVSM-RapCA. Biological replicates: 4 control replicates, 4 transduced replicates.

2011-04-13 | E-GEOD-24749 | biostudies-arrayexpress

Rap1a GTPase mVSM Knockout vs. Rescue mRNA Expression

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Rap1a GTPase mVSM Knockout vs. Rescue Expression

Project description:This SuperSeries is composed of the following subset Series: GSE24749: Rap1a GTPase mVSM Knockout vs. Rescue miRNA Expression GSE25702: Rap1a GTPase mVSM Knockout vs. Rescue mRNA Expression Refer to individual Series

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ARHGEF12 activates Rap1A and not RhoA in human capillary endothelial cells to reduce tumor necrosis factor-induced leak

Project description:In acute critical illness the capillary barrier may break down resulting in hypovolemia, extracellular edema, tissue dysoxia and even death. Restoration of the tight junction dependent capillary barrier is regulated by small GTPases, the specific regulatory molecules most active in this setting are not well described. Transcriptional profiling of tight junction forming human dermal microvascular ECs (HDMECs) and adherens junctions forming human umbilical vein EC (HUVECs) demonstrate ARHGEF12 is significantly differentially regulated in HDMECs before and after stimulation with tumor necrosis factor (TNF). HDMEC depleted of ArhGEF12 demonstrate significantly exacerbated TNF-induced decrease in trans-endothelial electrical resistance and disruption of tight junction proteins claudin-5 and ZO-1. ArhGEF12 is a RhoA-GEF, an established finding which is inconsistent with our observed results. Pulldown activation assays from of HDMECs depleted of ArhGEF12 and treated with TNF show decreased Rap1A activity after four hours and paradoxically increased RhoA activity after 12 hours. In cell-free pulldown assays, immunoprecipitated ArhGEF12 effectively activates Rap1A and RhoA and not Rap2A-C, RhoB-C or even Rap1B which shares 95% sequence identity with Rap1A. We conclude that in tight junction forming EC, ArhGEF12 selectively activates Rap1A to promote capillary barrier restoration in a mechanism independent of traditionally described cAMP-mediated Epac1 activation. The unique dual in vitro specificity of ArhGEF12 for RhoA, Rap1A and not Rap1B cannot be explained by sequence identity and requires additional investigation.

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Comparative proteomics of purified pathogen vacuoles correlates intracellular replication of Legionella pneumophila with the small GTPase Rap1

Project description:Legionella pneumophila is an opportunistic bacterial pathogen that causes a severe lung infection termed “Legionnaires’ disease”. The pathogen replicates in environmental protozoa as well as in macrophages within a unique membrane-bound compartment, the Legionella-containing-vacuole (LCV). Formation of the pathogen vacuole requires the bacterial Icm/Dot type IV secretion system (T4SS), which translocates ca. 300 effector proteins into host cells, where they subvert pivotal host processes and govern LCV formation. The L. pneumophila “pentuple” mutant lacks 5 gene clusters comprising 12% of the genome and 30% of the effector proteins. The mutant strain replicates in murine bone marrow-derived macrophages but not in Dictyostelium discoideum amoeba. In order to elucidate the host cell proteome defining a replication permissive compartment, we compare here the proteomes of intact LCVs isolated from D. discoideum or murine RAW 264.7 macrophages infected with the L. pneumophila parental strain Lp02 or the “pentuple” mutant. Proteome analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed host proteins specifically localizing to LCVs harbouring strain Lp02 or the “pentuple” mutant, respectively. The small GTPase Rap1 was identified on D. discoideum LCVs containing strain Lp02 but not the “pentuple” mutant and on macrophage LCVs containing either strain. The localization pattern of Rap1 on D. discoideum LCVs was confirmed by fluorescence microscopy, and depletion of Rap1 in A549 epithelial cells by RNA interference significantly reduced the intracellular growth of L. pneumophila. Thus, comparative proteomics identified Rap1 as a novel LCV host component implicated in intracellular replication of L. pneumophila.

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Mus musculus

Project description:Rap1a GTPase mVSM Knockout vs. Rescue miRNA Expression

| PRJNA142715 | ENA

Small GTPase Rap1A/B Is Required for Lymphatic Development and Adrenomedullin-Induced Stabilization of Lymphatic Endothelial Junctions.

Project description: Not available

| S-EPMC6206876 | biostudies-literature