Engineered Ribonucleoprotein Granules Inhibit Translation in Protocells

Simon, Joseph R.; Eghtesadi, Seyed Ali; Dzuricky, Michael; Li, You; Chilkoti, Ashutosh

doi:10.1016/j.molcel.2019.05.010

Cited by 60 publications

(61 citation statements)

References 52 publications

(62 reference statements)

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“…Additionally, they constitute a powerful tool for studying the structural properties that are responsible for phase transition of complex IDPs and the subsequent formation of protein-rich biomolecular condensates. [5][6][7][8] One of the most important families of IDPPs that has received increased attention in biomedical studies and applications are elastin-like recombinamers (ELRs). 4 These protein-engineered polymers are structurally based on the conserved motifs of tropoelastin, generally based on the pentapeptide with a sequence X1-Pro-Gly-X2-Gly (where X1 can be Val or Ile and X2 can be any amino acid except L-Pro).…”

Section: Introductionmentioning

confidence: 99%

Charge Density as a Molecular Modulator of Nanostructuration in Intrinsically Disordered Protein Polymers

et al. 2020

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Intrinsically disordered protein polymers (IDPPs) have attracted a lot of attention in the development of bioengineered devices and use as molecular biology study models due to their biomechanical properties and stimuli-responsiveness. The present work aims to understand the effect of charge distribution on self-assembly of IDPPs. To that end, a library of recombinant IDPPs based on an amphiphilic diblock design with different charge distributions were bioproduced and their supramolecular assembly characterized on the nano-, meso-and microscale. Although phase transition was driven by the collapse of hydrophobic moieties, hydrophilic block composition strongly affected hierarchical assembly and, therefore, enabled the production of new molecular

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Section: Introductionmentioning

confidence: 99%

Charge Density as a Molecular Modulator of Nanostructuration in Intrinsically Disordered Protein Polymers

et al. 2020

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“…Therefore, decoding the sequence determinants of intrinsically disordered protein (IDP) phase separation is important for understanding the biochemistry of biomolecular condensates in physiological and pathophysiological conditions. Characterizing the effects of sequence on phase behavior is also important for the field of protein-based materials (17), wherein proteins can be designed to have desired characteristics and programable assembly (18)(19)(20), with applications in biotechnology such as drug delivery, cell engineering, and biomimetics (21)(22)(23)(24).…”

mentioning

confidence: 99%

Identifying sequence perturbations to an intrinsically disordered protein that determine its phase-separation behavior

Schuster

Dignon

Tang

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

235

302

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Phase separation of intrinsically disordered proteins (IDPs) commonly underlies the formation of membraneless organelles, which compartmentalize molecules intracellularly in the absence of a lipid membrane. Identifying the protein sequence features responsible for IDP phase separation is critical for understanding physiological roles and pathological consequences of biomolecular condensation, as well as for harnessing phase separation for applications in bioinspired materials design. To expand our knowledge of sequence determinants of IDP phase separation, we characterized variants of the intrinsically disordered RGG domain from LAF-1, a model protein involved in phase separation and a key component of P granules. Based on a predictive coarse-grained IDP model, we identified a region of the RGG domain that has high contact probability and is highly conserved between species; deletion of this region significantly disrupts phase separation in vitro and in vivo. We determined the effects of charge patterning on phase behavior through sequence shuffling. We designed sequences with significantly increased phase separation propensity by shuffling the wild-type sequence, which contains well-mixed charged residues, to increase charge segregation. This result indicates the natural sequence is under negative selection to moderate this mode of interaction. We measured the contributions of tyrosine and arginine residues to phase separation experimentally through mutagenesis studies and computationally through direct interrogation of different modes of interaction using all-atom simulations. Finally, we show that despite these sequence perturbations, the RGG-derived condensates remain liquid-like. Together, these studies advance our fundamental understanding of key biophysical principles and sequence features important to phase separation.

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“…This mutation allowed the RGG domain to specifically recognize the RNA and was used to probe how TERRA regulates histone modifications (Takahama et al, ). Another more recent study used RGG domains fused to elastin‐like polypeptides to develop RBPs with tuneable phase behavior in protocells in an effort to study RBPs and RNA granules and showed that they granules can inhibit translation through either reversible or irreversible sequestration of mRNA (Simon, Eghtesadi, Dzuricky, You, & Chilkoti, ). Taken together these studies demonstrated that RGG boxes can be engineered for a variety of functions.…”

Section: Traditional Rbdsmentioning

confidence: 99%

The potential of engineered eukaryotic RNA‐binding proteins as molecular tools and therapeutics

2019

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Eukaroytic RNA‐binding proteins (RBPs) recognize and process RNAs through recognition of their sequence motifs via RNA‐binding domains (RBDs). RBPs usually consist of one or more RBDs and can include additional functional domains that modify or cleave RNA. Engineered RBPs have been used to answer basic biology questions, control gene expression, locate viral RNA in vivo, as well as many other tasks. Given the growing number of diseases associated with RNA and RBPs, engineered RBPs also have the potential to serve as therapeutics. This review provides an in depth description of recent advances in engineered RBPs and discusses opportunities and challenges in the field. This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein–RNA Recognition RNA Methods > RNA Nanotechnology RNA in Disease and Development > RNA in Disease

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Engineered Ribonucleoprotein Granules Inhibit Translation in Protocells

Cited by 60 publications

References 52 publications

Charge Density as a Molecular Modulator of Nanostructuration in Intrinsically Disordered Protein Polymers

Charge Density as a Molecular Modulator of Nanostructuration in Intrinsically Disordered Protein Polymers

Identifying sequence perturbations to an intrinsically disordered protein that determine its phase-separation behavior

The potential of engineered eukaryotic RNA‐binding proteins as molecular tools and therapeutics

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