Characterization of Membraneless Organelles
Compartmentalization is necessary for cells to carry out their physiological functions and maintain an organized environment. Traditionally, it was thought that organelles could only use membranes to achieve this physical separation, but there are many other organelles that achieve this separation without membranes, called membraneless organelles (MLOs). Biomolecular condensates include typical MLOs such as nucleoli, nuclear speckles, and stress granules, but also other components involved in transcription, translation, and signaling, such as heterochromatin, superenhancers, centrosomes, presynaptic and postsynaptic densities, and membrane receptor clusters. They exist as droplets within the cell and are formed by the condensation of cellular material in a process called liquid-liquid phase separation (LLPS). MLOs are numerous and diverse in the cell and support specific biochemistry with key functions in cellular homeostasis and development.
Fig. 1. Biomolecular condensates in eukaryotic cells. (Banani S F, et al., 2017)
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The compartments of MLOs are not confined by membranes, but still maintain a well-defined composition, structure and function. They are localized to different regions of the cell in bacteria and eukaryotes and play multiple important roles in a variety of cellular processes. Based on our cutting-edge LLPS platform and extensive experience, we integrate complementary approaches to help our clients study the structure, dynamics and biology of MLOs. We offer physical characterization of the following MLOs, including but not limited to:
| Cytoplasmic MLOs |
Stress granules, Processing bodies or P-bodies, GW bodies, Osmotic shock foci, Glucose starvation and Std1 cytoplasmic spots, Stock bodies, U-bodies, Neuronal RNA particles, Prion-induced ribonucleoprotein particles, Temporal asymmetry MRP bodies, Tight junctions and apical junctional condensates, Synapses, cGAS and cGAS-STING signaling, YBX1 condensates and microRNAs sorted into exosomes, Hyaline keratin granules, Cytoplasmic concentrates of MXA, GLC7 particles, DACT1 condensates and inhibition of Wnt signaling, Deup1 condensates, deuterostomes and multi-ciliogenesis, Kinetic protein axoneme particles, Ciliogenesis and BEX1 coagulation, RIα droplets, Smaug containing focal point, Synaptic XRN1 bodies, SA-induced condensation of NPR1, Viral factories. |
| Metabolic Condensates |
Glycolysis bodies, LLPS-accelerated H2B ubiquitination, Purinergic body. |
| Germ Cell MLOs |
RNP granules (grP bodies) in stalled or stressed oocytes, Germline P granules, Spongiosomes, Polar granules and polar plasma, Mitochondrial clouds, Germinal granules, Oskar vesicles in primordial germ cell, Chromatin bodies, Mouse testicular heat stress granules. |
| Organelles without Nuclear Membrane |
Nucleolus, Stages in the nucleosome stage, Nuclear pore complex, Parapodia, Nuclear speckles, Promyelocytic leukemia nucleosomes, Histone gene motifs, Caha bodies, Nuclear stress bodies, DNA damage foci, Sam68 nucleosomes, Dissociative bodies, DDX1 bodies, Nuclear gem or baryon of the coiled body, Enhancer clusters and phase separation of JMJD3, Oct1/PTF/transcriptional domain (OPT domain), Polymorphic interphase nucleosome association, GBPL Defense-activated cohesin, TopBP1 nuclear condensate, Proteasome foci, MORC3 nuclear condensate, STAT3 nucleosome, Phase separation in telomeric compartmentalization, NUP98-NSD1 fusion nuclear condensates, SMXL-containing subnuclear condensates, Nucleophosome, Viral nuclear droplets. |
| Mitochondrial and Chloroplast MLOs |
Mitochondrial RNA particles, Mitochondrial-like nuclei, RNA particles in chloroplasts, Chloroplast Stress Granules, Ribonucleic acid, Droplets and cargo sorting in chloroplasts. |
| Bacterial MLOs and Condensates |
BR bodies, Carboxysomes, Bacterial nucleoid, FtsZ condensates and bacterial division, SlmA and Noc clusters in bacterial nucleoid nuclei, Dps clusters, Bacterial RNA polymerase clusters, Compartmentalization complexes and ParABS DNA separation systems, Compartmentalized ABC translocator proteins, Bacterial cell poles, Single-stranded DNA-binding droplets, PlyP particles, Plec-PodJ condensate. |
As your ideal partner, CD BioSciences offers comprehensive approaches to achieve physical characterization of the structure and dynamics of MLOs and their internal molecules on picosecond to hour time scales and Å to millimeter length scales, including:
- Morphological Characterization of MLOs
We have an extensive microscopy platform to perform detailed morphological characterization of biomolecular condensates.
- Optical microscopy allows visualization of biomolecules in vitro and in living cells.
- Super-resolution microscopy combines optical input with mathematical analysis to analyze relevant changes in the morphology of biomolecular condensates.
- Electron microscopy can be used to separate components of molecular and macromolecular phases formed in vitro, such as hydrogels and protofibrils.
- Rheology of MLOs
We can help you analyze the material properties of membraneless organelles (e.g., viscosity, surface tension, molecular network mesh size, etc.) as well as the dynamics of the constituent molecules.
- Characterization of MLOs Composition
Most MLOs are composed of proteins and nucleic acids. We used unbiased large-scale approaches (proteomics and transcriptomics) to analyze the biomolecular composition of a variety of different MLOs.
- Structural Features of MLOs
CD BioSciences has undergone a multidisciplinary effort to combine multiple approaches to help clients analyze the composition, structure, and dynamic characteristics of MLOs. In addition, our experts are developing innovative approaches to address the complex structures and functions associated with MLOs. Our services can provide useful guidance for future scientists to discover how biological structures formed by MLOs through phase separation enable molecular processes critical to life. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together with your attractive projects.
Reference
- Banani S F, Lee H O, Hyman A A, et al. (2017) Biomolecular condensates: organizers of cellular biochemistry[J]. Nature reviews Molecular cell biology. 18(5): 285-298.
For research use only, not intended for any clinical use.
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