Articles in 2022

Phase separation is an important mechanism for biomolecule condensate assembly and is involved in multiple biological activities. Understanding its molecular mechanism provides a unique perspective for gaining insights into its role in cellular physiology and for developing new tools for the manipulation of cellular function.

This perspective proposes general strategies for phase-separation-related biological studies, including proper experimental designs to validate and characterize phase-separation phenomena, connections to biological functions and some caveats to avoid common misunderstandings.

Protein condensates are subcellular structures that enrich and confine molecules in cells. This Review details how condensates can be engineered with responsiveness and on-demand functions, thus pushing cellular and metabolic engineering to a new level.

This Review introduces molecular features of the phase-separating biomolecules and how they affect phase-separation behavior in a complex intracellular environment, highlighting a complex interplay between structure, sequence and environment in the phase-separation process.

Clustering and multimerization of cell surface proteins (CSPs) are essential for triggering downstream intracellular signaling events. Membrane-anchored liquid–liquid phase-separation systems have now been developed to manipulate the spatiotemporal distribution and activation of CSPs.

A membrane-tethering liquid–liquid phase-separation system was developed for programmable compartmentalization of cell-surface proteins and regulation of downstream cellular activities.

SEUSS is a transcriptional adaptor that undergoes condensation after hyperosmotic stress-induced increase of molecular crowding. The SEUSS condensates are indispensable for stress tolerance via facilitating the expression of stress-related genes.

The non-nucleoside reverse transcriptase inhibitors (NNRTIs) drive HIV protease-mediated CARD8 inflammasome activation, which is attenuated by dipeptidyl peptidase 9 (DPP9). DPP9 inhibitors act synergistically with NNRTIs to clear HIV-infected cells.

Coupling haploid genetics with deep scanning mutagenesis, Hanzl et al. identified functional hotspots in E3 ubiquitin ligases that are selectively required for different proteolysis-targeting chimeras (PROTACs) or molecular glue degraders and found mutated in relapsing patients.

Akizuki et al. reveal an unexpected role for K63-linked ubiquitin chains and the E2 enzyme UBE2N in degrader-induced degradation of cIAP1 through the proteasome, demonstrating the diversity of the ubiquitin code used for targeted degradation.

Coenzyme A (CoA) is a ubiquitous and essential cofactor. A biosensor for visualizing cytosolic and mitochondrial CoA in living cells was developed to address central questions concerning CoA homeostasis.

Velcrins kill cancer cells by inducing complex formation between PDE3A and SLFN12, upregulating SLFN12 RNase activity. Activated SLFN12 specifically cleaves tRNA^Leu(TAA), resulting in global inhibition of protein synthesis.

Fan et al. report a potent and subtype-selective TRPV3 antagonist, Trpvicin, and reveal its binding sites and mode of action for TRPV3 inhibition via high-resolution cryogenic electron microscopy structures.

The cryo-EM structures of MRGPRX1–Gq complexes are reported, which revealed the activation and allosteric modulation mechanism of human MRGPRX1 receptor, which may enable the structure-based identification of novel analgesics.

Murray et al. identified and characterized a small-molecule inhibitor of human COQ8A, which belongs to the UbiB protein family and is essential for coenzyme Q biosynthesis.

YcaO enzymes are able to catalyze a diverse set of reactions and have found industrial applications. New biochemical data provide the first direct evidence for the unified reaction mechanism proposed a decade ago and will inform future enzyme engineering efforts.