Dataset Information

Optically inducible membrane recruitment and signaling systems.

ABSTRACT: Optical induction of intracellular signaling by membrane-associated and integral membrane proteins allows spatiotemporally precise control over second messenger signaling and cytoskeletal rearrangements that are important to cell migration, development, and proliferation. Optogenetic membrane recruitment of a protein-of-interest to control its signaling by altering subcellular localization is a versatile means to these ends. Here, we summarize the signaling characteristics and underlying structure-function of RGS-LOV photoreceptors as single-component membrane recruitment tools that rapidly, reversibly, and efficiently carry protein cargo from the cytoplasm to the plasma membrane by a light-regulated electrostatic interaction with the membrane itself. We place the technology-relevant features of these recently described natural photosensory proteins in context of summarized protein engineering and design strategies for optically controlling membrane protein signaling.

SUBMITTER: Hannanta-Anan P

PROVIDER: S-EPMC6697567 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Optically inducible membrane recruitment and signaling systems.

Hannanta-Anan Pimkhuan P Glantz Spencer T ST Chow Brian Y BY

Current opinion in structural biology 20190316

Optical induction of intracellular signaling by membrane-associated and integral membrane proteins allows spatiotemporally precise control over second messenger signaling and cytoskeletal rearrangements that are important to cell migration, development, and proliferation. Optogenetic membrane recruitment of a protein-of-interest to control its signaling by altering subcellular localization is a versatile means to these ends. Here, we summarize the signaling characteristics and underlying structu ...[more]

PMID: 30884362

Similar Datasets

Project description:Cell membrane capacitance and conductance are key pieces of intrinsic information correlated with the cellular dielectric parameters and morphology of the plasma membrane; these parameters have been used as electrophysiological biomarkers to characterize cellular phenotype and state, and they have many associated clinical applications. Here, we present our work on the non-invasive determination of cell membrane capacitance and conductance by an optically activated microfluidics chip. The model for determining the cell membrane capacitance and conductance was established by a single layer of the shell-core polarization model. Three-dimensional finite-element analyses of the positive and negative optically induced dielectrophoresis forces generated by the projected light arrays of spots were performed, thus providing a theoretical validation of the feasibility of this approach. Then, the crossover frequency spectra for four typical types of cells (Raji cells, MCF-7 cells, HEK293 cells, and K562 cells) were experimentally investigated by using a micro-vision based motion-tracking technique. The different responses of these cells to the positive and negative ODEP forces were studied under four different liquid conductivities by automatic observation and tracking of the cellular trajectory and texture during the cells' translation. The cell membrane capacitance and conductance were determined from the curve-fitted spectra, which were 11.1 ± 0.9 mF/m2 and 782 ± 32 S/m2, respectively, for Raji cells, 11.5 ± 0.8 mF/m2 and 114 ± 28 S/m2 for MCF-7 cells, 9.0 ± 0.9 mF/m2 and 187 ± 22 S/m2 for HEK293 cells, and 10.2 ± 0.7 mF/m2 and 879 ± 24 S/m2 for K562 cells. Furthermore, as an application of this technique, the membrane capacitances of MCF-7 cells treated with four different concentrations of drugs were acquired. This technique introduces a determination of cell membrane capacitance and conductance that yields statistically significant data while allowing information from individual cells to be obtained in a non-invasive manner.

Project description:Cysteine donates sulfur to macromolecules and occurs naturally in many proteins. Because low concentrations of cysteine are cytotoxic, its intracellular concentration is stringently controlled. In bacteria, cysteine biosynthesis is regulated by feedback inhibition of the activities of serine acetyltransferase (SAT) and 3-phosphoglycerate dehydrogenase (3-PGDH) and is also regulated at the transcriptional level by inducing the cysteine regulon using the master regulator CysB. Here, we describe two novel cysteine-inducible systems that regulate the cysteine resistance of Pantoea ananatis, a member of the family Enterobacteriaceae that shows great potential for producing substances useful for biotechnological, medical, and industrial purposes. One locus, designated ccdA(formerly PAJ_0331), encodes a novel cysteine-inducible cysteine desulfhydrase (CD) that degrades cysteine, and its expression is controlled by the transcriptional regulator encoded byccdR(formerly PAJ_0332 orybaO), located just upstream of ccdA The other locus, designated cefA (formerly PAJ_3026), encodes a novel cysteine-inducible cysteine efflux pump that is controlled by the transcriptional regulator cefR(formerly PAJ_3027), located just upstream of cefA To our knowledge, this is the first example where the expression of CD and an efflux pump is regulated in response to cysteine and is directly involved in imparting resistance to excess levels of cysteine. We propose that ccdA and cefA function as safety valves that maintain homeostasis when the intra- or extracellular cysteine concentration fluctuates. Our findings contribute important insights into optimizing the production of cysteine and related biomaterials by P. ananatisBecause of its toxicity, the bacterial intracellular cysteine level is stringently regulated at biosynthesis. This work describes the identification and characterization of two novel cysteine-inducible systems that regulate, through degradation and efflux, the cysteine resistance of Pantoea ananatis, a member of the family Enterobacteriaceae that shows great potential for producing substances useful for industrial purposes. We propose that this novel mechanism for sensing and regulating cysteine levels is a safety valve enabling adaptation to sudden changes in intra- or extracellular cysteine levels in bacteria. Our findings provide important insights into optimizing the production of cysteine and related biomaterials by P. ananatis and also a deep understanding of sulfur/cysteine metabolism and regulation in this plant pathogen and related bacteria.

Dataset Information

Optically inducible membrane recruitment and signaling systems.

Publications

Optically inducible membrane recruitment and signaling systems.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure