Salmonella exploits membrane reservoirs for invasion of host cells
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ABSTRACT: Submitted data contain the results of BioID-LC-MS experiments performed in T-REx HEK293 expressing RAB10 protein (WT and mutants) fused to BirA biotin carboxylase.
Project description:Data deposited contain results from proximity-dependent biotinylation LC-MS (BioID) experiments in which T-REx Flp-In HEK293 cells express SCARB1 protein fused to biotin carboxylase BirA.
Project description:Data deposited contain results from proximity-dependent biotinylation LC-MS (BioID) experiments in which T-REx Flp-In HEK293 cells express ClpX protein fused to miniTurbo biotin carboxylase with or without a mitochondrial localization signal.
Project description:Submission contains data from proximity labeling combined to mass spectrometry (BioID) with protein of interest TPMT fused in-frame with BirA-R118G carboxylase expressed in HEK293 T-REx Flp-In cells. Proximity interacting proteins were captured with streptavidin sepharose, digested with trypsin and analyzed by LC-MS.
Project description:Data deposited contain results from proximity-dependent biotinylation LC-MS (BioID) experiments in which T-REx Flp-In HEK293 cells express NSD2 protein fused to miniTurbo biotin carboxylase with or without treatment with the protac UNC8732.
Project description:Salmonella utilizes a type 3 secretion system to translocate virulence proteins (effectors) into host cells during infection1. The effectors modulate host cell machinery to drive uptake of the bacteria into vacuoles, where they can establish an intracellular replicative niche. A remarkable feature of Salmonella invasion is the formation of actin-rich protuberances (ruffles) on the host cell surface that contribute to bacterial uptake. However, the membrane source for ruffle formation and how these bacteria regulate membrane mobilization within host cells remains unclear. Here, we show that Salmonella exploits membrane reservoirs for the generation of invasion ruffles. The reservoirs are pre-existing tubular compartments associated with the plasma membrane (PM) and are formed through the activity of RAB10 GTPase. Under normal growth conditions, membrane reservoirs contribute to PM homeostasis and are preloaded with the exocyst subunit EXOC2. During Salmonella invasion, the bacterial effectors SipC, SopE2, and SopB recruit exocyst subunits from membrane reservoirs and other cellular compartments, thereby allowing exocyst complex assembly and membrane delivery required for bacterial uptake. Our findings reveal an important role for RAB10 in the establishment of membrane reservoirs and the mechanisms by which Salmonella can exploit these compartments during host cell invasion.
Project description:Data deposited contain results from proximity-dependent biotinylation LC-MS (BioID) experiments in which T-REx Flp-In HEK293, 697 and HAP1 cells expressing the E3 ligase VHL protein fused to miniTurbo biotin carboxylase were treated with either DMSO or various protacs with or without proteasome inhibitors.
Project description:Here we investigated the effects of CEBPA transcription factor expression on myeloid NB4 cells. The sequence of rat CEBPA was C-terminally fused to a promiscuous biotin ligase tag (BirA*) and NB4 cell lines were engineered to express the fusion protein under the control of a doxycycline inducible promoter. Three different NB4 cell lines were investigated that expressed (i) BirA* tag alone (ii) full length CEBPA isoform (P42) fused to BirA* (iii) truncated CEBPA isoform (P30) fused to BirA*. Cells were seeded in media supplemented with or without doxycycline.
Project description:Data contains LC-MS data of anti-Flag affinity purification to generate the protein interaction network of SopD protein fused with BirAFlag (C-terminal tag)
and expressed in T-REx HEK293 cells.
Project description:We report the genomic localization of cohesin oligomers in nocodazole arrested yeast cells. Two alleles of SMC3 were expressed in yeast cells, one fused to BirA enzyme and the other tagged with AviTag. Cohesin oligomers were biotinylated and ChIP with streptavidin beads. As control experiments, cohesin localization on chromosome was determined in strains expresses freely diffusable BirA enzyme, where all Smc3 proteins were biotinylated; non-specific ChIP were determined in strains with no BirA.