Dataset Information

LITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution.

ABSTRACT: Many bacteria employ a type III secretion system (T3SS) injectisome to translocate proteins into eukaryotic host cells. Although the T3SS can efficiently export heterologous cargo proteins, a lack of target cell specificity currently limits its application in biotechnology and healthcare. In this study, we exploit the dynamic nature of the T3SS to govern its activity. Using optogenetic interaction switches to control the availability of the dynamic cytosolic T3SS component SctQ, T3SS-dependent effector secretion can be regulated by light. The resulting system, LITESEC-T3SS (Light-induced translocation of effectors through sequestration of endogenous components of the T3SS), allows rapid, specific, and reversible activation or deactivation of the T3SS upon illumination. We demonstrate the light-regulated translocation of heterologous reporter proteins, and induction of apoptosis in cultured eukaryotic cells. LITESEC-T3SS constitutes a new method to control protein secretion and translocation into eukaryotic host cells with unparalleled spatial and temporal resolution.

SUBMITTER: Lindner F

PROVIDER: S-EPMC7221075 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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LITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution.

Lindner Florian F Milne-Davies Bailey B Langenfeld Katja K Stiewe Thorsten T Diepold Andreas A

Nature communications 20200513 1

Many bacteria employ a type III secretion system (T3SS) injectisome to translocate proteins into eukaryotic host cells. Although the T3SS can efficiently export heterologous cargo proteins, a lack of target cell specificity currently limits its application in biotechnology and healthcare. In this study, we exploit the dynamic nature of the T3SS to govern its activity. Using optogenetic interaction switches to control the availability of the dynamic cytosolic T3SS component SctQ, T3SS-dependent e ...[more]

PMID: 32404906

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Project description:Most studies on the biological effects of future climatic changes rely on seasonally aggregated, coarse-resolution data. Such data mask spatial and temporal variability in microclimate driven by terrain, wind and vegetation, and ultimately bear little resemblance to the conditions that organisms experience in the wild. Here, I present the methods for providing fine-grained, hourly and daily estimates of current and future temperature and soil moisture over decadal timescales. Observed climate data and spatially coherent probabilistic projections of daily future weather were disaggregated to hourly and used to drive empirically calibrated physical models of thermal and hydrological microclimates. Mesoclimatic effects (cold-air drainage, coastal exposure and elevation) were determined from the coarse-resolution climate surfaces using thin-plate spline models with coastal exposure and elevation as predictors. Differences between micro and mesoclimate temperatures were determined from terrain, vegetation and ground properties using energy balance equations. Soil moisture was computed in a thin upper layer and an underlying deeper layer, and the exchange of water between these layers was calculated using the van Genuchten equation. Code for processing the data and running the models is provided as a series of R packages. The methods were applied to the Lizard Peninsula, United Kingdom, to provide hourly estimates of temperature (100 m grid resolution over entire area, 1 m for a selected area) for the periods 1983-2017 and 2041-2049. Results indicated that there is a fine-resolution variability in climatic changes, driven primarily by interactions between landscape features and decadal trends in weather conditions. High-temporal resolution extremes in conditions under future climate change were predicted to be considerably less novel than the extremes estimated using seasonally aggregated variables. The study highlights the need to more accurately estimate the future climatic conditions experienced by organisms and equips biologists with the means to do so.

Project description:PurposeTo prospectively evaluate the feasibility of performing high-spatial-resolution (1-mm isotropic) time-resolved three-dimensional (3D) contrast material-enhanced magnetic resonance (MR) angiography of the peripheral vasculature with Cartesian acquisition with projection-reconstruction-like sampling (CAPR) and eightfold accelerated two-dimensional (2D) sensitivity encoding (SENSE).Materials and methodsAll studies were approved by the institutional review board and were HIPAA compliant; written informed consent was obtained from all participants. There were 13 volunteers (mean age, 41.9; range, 27-53 years). The CAPR sequence was adapted to provide 1-mm isotropic spatial resolution and a 5-second frame time. Use of different receiver coil element sizes for those placed on the anterior-to-posterior versus left-to-right sides of the field of view reduced signal-to-noise ratio loss due to acceleration. Results from eight volunteers were rated independently by two radiologists according to prominence of artifact, arterial to venous separation, vessel sharpness, continuity of arterial signal intensity in major arteries (anterior and posterior tibial, peroneal), demarcation of origin of major arteries, and overall diagnostic image quality. MR angiographic results in two patients with peripheral vascular disease were compared with their results at computed tomographic angiography.ResultsThe sequence exhibited no image artifact adversely affecting diagnostic image quality. Temporal resolution was evaluated to be sufficient in all cases, even with known rapid arterial to venous transit. The vessels were graded to have excellent sharpness, continuity, and demarcation of the origins of the major arteries. Distal muscular branches and the communicating and perforating arteries were routinely seen. Excellent diagnostic quality rating was given for 15 (94%) of 16 evaluations.ConclusionThe feasibility of performing high-diagnostic-quality time-resolved 3D contrast-enhanced MR angiography of the peripheral vasculature by using CAPR and eightfold accelerated 2D SENSE has been demonstrated.

Dataset Information

LITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution.

Publications

LITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution.

Similar Datasets

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