Structure of Semliki Forest virus in complex with its receptor VLDLR.

Semliki Forest virus (SFV) is an alphavirus that uses the very-low-density lipoprotein receptor (VLDLR) as a receptor during infection of its vertebrate hosts and insect vectors. Herein, we used cryoelectron microscopy to study the structure of SFV in complex with VLDLR. We found that VLDLR binds multiple E1-DIII sites of SFV through its membrane-distal LDLR class A (LA) repeats. Among the LA repeats of the VLDLR, LA3 has the best binding affinity to SFV. The high-resolution structure shows that LA3 binds SFV E1-DIII through a small surface area of 378 Å2, with the main interactions at the interface involving salt bridges. Compared with the binding of single LA3s, consecutive LA repeats around LA3 promote synergistic binding to SFV, during which the LAs undergo a rotation, allowing simultaneous key interactions at multiple E1-DIII sites on the virion and enabling the binding of VLDLRs from divergent host species to SFV.

Structure Studies of the CRISPR-Csm Complex Reveal Mechanism of Co-transcriptional Interference.

Csm, a type III-A CRISPR-Cas interference complex, is a CRISPR RNA (crRNA)-guided RNase that also possesses target RNA-dependent DNase and cyclic oligoadenylate (cOA) synthetase activities. However, the structural features allowing target RNA-binding-dependent activation of DNA cleavage and cOA generation remain unknown. Here, we report the structure of Csm in complex with crRNA together with structures of cognate or non-cognate target RNA bound Csm complexes. We show that depending on complementarity with the 5' tag of crRNA, the 3' anti-tag region of target RNA binds at two distinct sites of the Csm complex. Importantly, the interaction between the non-complementary anti-tag region of cognate target RNA and Csm1 induces a conformational change at the Csm1 subunit that allosterically activates DNA cleavage and cOA generation. Together, our structural studies provide crucial insights into the mechanistic processes required for crRNA-meditated sequence-specific RNA cleavage, RNA target-dependent non-specific DNA cleavage, and cOA generation.

The Molecular Architecture for RNA-Guided RNA Cleavage by Cas13a.

Cas13a, a type VI-A CRISPR-Cas RNA-guided RNA ribonuclease, degrades invasive RNAs targeted by CRISPR RNA (crRNA) and has potential applications in RNA technology. To understand how Cas13a is activated to cleave RNA, we have determined the crystal structure of Leptotrichia buccalis (Lbu) Cas13a bound to crRNA and its target RNA, as well as the cryo-EM structure of the LbuCas13a-crRNA complex. The crRNA-target RNA duplex binds in a positively charged central channel of the nuclease (NUC) lobe, and Cas13a protein and crRNA undergo a significant conformational change upon target RNA binding. The guide-target RNA duplex formation triggers HEPN1 domain to move toward HEPN2 domain, activating the HEPN catalytic site of Cas13a protein, which subsequently cleaves both single-stranded target and collateral RNAs in a non-specific manner. These findings reveal how Cas13a of type VI CRISPR-Cas systems defend against RNA phages and set the stage for its development as a tool for RNA manipulation.

In situ structure of the red algal phycobilisome-PSII-PSI-LHC megacomplex.

In oxygenic photosynthetic organisms, light energy is captured by antenna systems and transferred to photosystem II (PSII) and photosystem I (PSI) to drive photosynthesis1,2. The antenna systems of red algae consist of soluble phycobilisomes (PBSs) and transmembrane light-harvesting complexes (LHCs)3. Excitation energy transfer pathways from PBS to photosystems remain unclear owing to the lack of structural information. Here we present in situ structures of PBS-PSII-PSI-LHC megacomplexes from the red alga Porphyridium purpureum at near-atomic resolution using cryogenic electron tomography and in situ single-particle analysis4, providing interaction details between PBS, PSII and PSI. The structures reveal several unidentified and incomplete proteins and their roles in the assembly of the megacomplex, as well as a huge and sophisticated pigment network. This work provides a solid structural basis for unravelling the mechanisms of PBS-PSII-PSI-LHC megacomplex assembly, efficient energy transfer from PBS to the two photosystems, and regulation of energy distribution between PSII and PSI.

Basis of the H2AK119 specificity of the Polycomb repressive deubiquitinase.

Repression of gene expression by protein complexes of the Polycomb group is a fundamental mechanism that governs embryonic development and cell-type specification1-3. The Polycomb repressive deubiquitinase (PR-DUB) complex removes the ubiquitin moiety from monoubiquitinated histone H2A K119 (H2AK119ub1) on the nucleosome4, counteracting the ubiquitin E3 ligase activity of Polycomb repressive complex 1 (PRC1)5 to facilitate the correct silencing of genes by Polycomb proteins and safeguard active genes from inadvertent silencing by PRC1 (refs. 6-9). The intricate biological function of PR-DUB requires accurate targeting of H2AK119ub1, but PR-DUB can deubiquitinate monoubiquitinated free histones and peptide substrates indiscriminately; the basis for its exquisite nucleosome-dependent substrate specificity therefore remains unclear. Here we report the cryo-electron microscopy structure of human PR-DUB, composed of BAP1 and ASXL1, in complex with the chromatosome. We find that ASXL1 directs the binding of the positively charged C-terminal extension of BAP1 to nucleosomal DNA and histones H3-H4 near the dyad, an addition to its role in forming the ubiquitin-binding cleft. Furthermore, a conserved loop segment of the catalytic domain of BAP1 is situated near the H2A-H2B acidic patch. This distinct nucleosome-binding mode displaces the C-terminal tail of H2A from the nucleosome surface, and endows PR-DUB with the specificity for H2AK119ub1.

Structure of Venezuelan equine encephalitis virus with its receptor LDLRAD3.

Venezuelan equine encephalitis virus (VEEV) is an enveloped RNA virus that causes encephalitis and potentially mortality in infected humans and equines1. At present, no vaccines or drugs are available that prevent or cure diseases caused by VEEV. Low-density lipoprotein receptor class A domain-containing 3 (LDLRAD3) was recently identified as a receptor for the entry of VEEV into host cells2. Here we present the cryo-electron microscopy structure of the LDLRAD3 extracellular domain 1 (LDLRAD3-D1) in complex with VEEV virus-like particles at a resolution of 3.0 Å. LDLRAD3-D1 has a cork-like structure and is inserted into clefts formed between adjacent VEEV E2-E1 heterodimers in the viral-surface trimer spikes through hydrophobic and polar contacts. Mutagenesis studies of LDLRAD3-D1 identified residues that are involved in the key interactions with VEEV. Of note, some of the LDLRAD3-D1 mutants showed a significantly increased binding affinity for VEEV, suggesting that LDLRAD3-D1 may serve as a potential scaffold for the development of inhibitors of VEEV entry. Our structures provide insights into alphavirus assembly and the binding of receptors to alphaviruses, which may guide the development of therapeutic countermeasures against alphaviruses.

Cryo-EM structures of LolCDE reveal the molecular mechanism of bacterial lipoprotein sorting in Escherichia coli.

Bacterial lipoproteins perform a diverse array of functions including bacterial envelope biogenesis and microbe-host interactions. Lipoproteins in gram-negative bacteria are sorted to the outer membrane (OM) via the localization of lipoproteins (Lol) export pathway. The ATP-binding cassette (ABC) transporter LolCDE initiates the Lol pathway by selectively extracting and transporting lipoproteins for trafficking. Here, we report cryo-EM structures of LolCDE in apo, lipoprotein-bound, and AMPPNP-bound states at a resolution of 3.5 to 4.2 Å. Structure-based disulfide crosslinking, photo-crosslinking, and functional complementation assay verify the apo-state structure and reveal the molecular details regarding substrate selectivity and substrate entry route. Our studies snapshot 3 functional states of LolCDE in a transport cycle, providing deep insights into the mechanisms that underlie LolCDE-mediated lipoprotein sorting in E. coli.

Bulk photovoltaic effect in ferroelectric nematic liquid crystals.

The bulk photovoltaic (BPV) effect in ferroelectric liquid crystals is of increasing scientific interest owing to its great potential for light-energy conversion. The ferroelectric nematic phase exhibits a huge spontaneous polarization that can be aligned to a preferred direction. In this Letter, we investigate the tensorial properties of the BPV effect in the planarly aligned ferroelectric nematic phase of the liquid crystalline material RM734. A steady-state short-circuit photocurrent of ~160 pA and an open-circuit photovoltage of ~50 mV were observed in a cell with a thickness of 5.5 µm under the illumination of ultraviolet light without any bias voltage. Based on the photocurrent measurements in different electrode configurations, the non-zero elements of the BPV tensor were obtained. The BPV effect is attributed to the combination of the spontaneous polarization and the asymmetric distribution of photoinduced charge carriers. This study not only provides an understanding of the bulk PV mechanism in soft ferroelectrics but also promises a wide range of unprecedented, to the best of our knowledge, benefits for light harvesting to engineer marketable photovoltaic devices.

Thermally tunable anti-ambipolar heterojunction devices.

Two-dimensional materials and their van der Waals heterostructures have emerged as a research focal point for constructing various innovative electronic devices due to their distinct photonic and electronic properties. Among them, anti-ambipolar devices, characterized by their unique nonlinear electrical behavior, have garnered attention as novel multifunctional components, positioning them as potential contenders for building multi-state logic devices. Utilizing the properties of few-layer As0.4P0.6 and PdSe2, we have constructed an anti-ambipolar heterojunction device. At 300 K, the device exhibits a peak voltage (Vpeak) of -3 V and a peak-to-valley ratio (PVR) close to 8 × 103, and the PVR can be modulated by bias voltage. Furthermore, by characterizing the anti-ambipolar attributes at different temperatures ranging from 80 K to 330 K, we have elucidated the thermally tunable feature of the device. At 330 K, a certain PVR (∼103) and a large Vpeak (∼-16 V) are obtained, while a PVR exceeding 108 has been achieved at 80 K. This temperature-related sensitivity empowers the device with significant potential and thermal tunability in various applications.

Insights into current bio-processes and future perspectives of carbon-neutral treatment of industrial organic wastewater: A critical review.

With the rise of the concept of carbon neutrality, the current wastewater treatment process of industrial organic wastewater is moving towards the goal of energy conservation and carbon emission reduction. The advantages of anaerobic digestion (AD) processes in industrial organic wastewater treatment for bio-energy recovery, which is in line with the concept of carbon neutrality. This study summarized the significance and advantages of the state-of-the-art AD processes were reviewed in detail. The application of expanded granular sludge bed (EGSB) reactors and anaerobic membrane bioreactor (AnMBR) were particularly introduced for the effective treatment of industrial organic wastewater treatment due to its remarkable prospect of engineering application for the high-strength wastewater. This study also looks forward to the optimization of the AD processes through the enhancement strategies of micro-aeration pretreatment, acidic-alkaline pretreatment, co-digestion, and biochar addition to improve the stability of the AD system and energy recovery from of industrial organic wastewater. The integration of anaerobic ammonia oxidation (Anammox) with the AD processes for the post-treatment of nitrogenous pollutants for the industrial organic wastewater is also introduced as a feasible carbon-neutral process. The combination of AnMBR and Anammox is highly recommended as a promising carbon-neutral process for the removal of both organic and inorganic pollutants from the industrial organic wastewater for future perspective. It is also suggested that the AD processes combined with biological hydrogen production, microalgae culture, bioelectrochemical technology and other bio-processes are suitable for the low-carbon treatment of industrial organic wastewater with the concept of carbon neutrality in future.

Insights into carbon-neutral treatment of rural wastewater by constructed wetlands: A review of current development and future direction.

In recent years, with the global rise in awareness regarding carbon neutrality, the treatment of wastewater in rural areas is increasingly oriented towards energy conservation, emission reduction, low-carbon output, and resource utilization. This paper provides an analysis of the advantages and disadvantages of the current low-carbon treatment process of low-carbon treatment for rural wastewater. Constructed wetlands (CWs) are increasingly being considered as a viable option for treating wastewater in rural regions. In pursuit of carbon neutrality, advanced carbon-neutral bioprocesses are regarded as the prospective trajectory for achieving carbon-neutral treatment of rural wastewater. The incorporation of CWs with emerging biotechnologies such as sulfur-based autotrophic denitrification (SAD), pyrite-based autotrophic denitrification (PAD), and anaerobic ammonia oxidation (anammox) enables efficient removal of nitrogen and phosphorus from rural wastewater. The advancement of CWs towards improved removal of organic and inorganic pollutants, sustainability, minimal energy consumption, and low carbon emissions is widely recognized as a viable low-carbon approach for achieving carbon-neutral treatment of rural wastewater. This study offers novel perspectives on the sustainable development of wastewater treatment in rural areas within the framework of achieving carbon neutrality in the future.

Cryo-EM structure of the nonameric CsgG-CsgF complex and its implications for controlling curli biogenesis in Enterobacteriaceae.

Curli play critical roles in biofilm formation, host cell adhesion, and colonization of inert surfaces in many Enterobacteriaceae. In Escherichia coli, curli biogenesis requires 7 curli-specific gene (csg) products-CsgA through G-working in concert. Of them, CsgG and CsgF are 2 outer membrane (OM)-localized components that consists of the core apparatus for secretion and assembly of curli structural subunits, CsgB and CsgA. Here, we report the cryogenic electron microscopy (cryo-EM) structure of CsgG in complex with CsgF from E. coli. The structure reveals that CsgF forms a stable complex with CsgG via a 1:1 stoichiometry by lining the upper lumen of the nonameric CsgG channel via its N-terminal 27 residues, forming a funnel-like entity plugged in the CsgG channel and creating a unique secretion channel with 2 constriction regions, consistent with the recently reported structure of the CsgG-CsgF complex. Functional studies indicate that export of CsgF to the cell surface requires the CsgG channel, and CsgF not only functions as an adaptor that bridges CsgB with CsgG but also may play important roles in controlling the rates of translocation and/or polymerization for curli structural subunits. Importantly, we found that a series of CsgF-derived peptides are able to efficiently inhibit curli production to E. coli when administrated exogenously, highlighting a potential strategy to interfere biofilm formation in E. coli strains.

Purification-induced damage to calicivirus particles at near-atomic resolution.

The purification of virus particles is an essential process for the manufacture of vaccines. However, the application of different purification processes may affect the quality of the virus particles, such as structural integrity and homogeneity, which may further influence the infectivity and immunogenicity of the purified virus. In this study, we took Feline calicivirus (FCV), a common natural pathogen in cats belonging to Caliciviridae, as a research model. By using cryo-electron microscopy (cryo-EM), we incorporated the 3D classification process as a virus flexibility evaluation system. Cryo-EM images of virus particles resulting from different purification processes were compared at near-atomic resolution. The results indicated that molecular sieving purification will impact the stability of P-domains through increasing flexibility as determined by the evaluation system, which can be extended to assess the purification effect on the entire particle. This evaluation process can be further applied to all non-enveloped viruses.

Interface characteristics of graphene/ZnS hybrid-dimensional heterostructures.

Graphene/ZnS hybrid-dimensional heterostructure is an excellent combination to regulate and improve the conductivity and sensitivity of components, in which the interface effects have crucial impacts on the performance of devices. In this work, we investigate the interface characteristics of Graphene/ZnS 2D/3D heterostructures. X-ray photoelectron spectra show that the ZnS binding energy shifts to lower energy by 0.3 eV after forming heterojunction with graphene. The fluorescence and absorption spectra confirm the luminescence enhancement and blue-shift of the absorbance edge of ZnS caused by graphene. The composition of Graphene/ZnS heterostructure facilitates separation and transfer of spatial charges, resulting in rapid electron transport.

Plasmon-induced transparency in a reconfigurable composite valley photonic crystal.

We propose a new kind of reconfigurable topological valley photonic crystal (TVPC), and a novel topological waveguide can be formed by constructing a domain wall between two TVPCs with opposite valley-Chern indices. The topological waveguide mode in the composite TVPC has large group refractive index. A topologically protected coupled waveguide cavity system is then designed by introducing a hexagonal ring cavity at the center of the straight domain wall of a combined TVPC, in which a narrow plasmon induced transparency window rises at 3.8848 GHz with a Q-factor of 1387 and a maximum group refractive index as high as 186. We propose a notch filter with a resonant frequency of 3.8852 GHz and a very high Q-factor of 10224. By changing the refractive index of liquid crystals via an external voltage applied between two parallel metal plates, the filter can be switched between band-pass and band-stop based on the reconfigurable topological interface state.

Metasurfaces with high-Q resonances governed by topological edge state.

Achieving high-quality (Q)-factor resonances in metasurfaces is essential for various applications, including nano-lasers, nonlinear optics, and quantum optics. In this work, we propose a high-Q metasurface using a topological strategy: constructing the metasurface by stacking two conjugated nanopillar arrays with different topological invariants. Our study shows that a topological edge state steadily appears at the interfaces of the nanopillars, and a sharp transmission resonance with a Q-factor of more than 1000 can be obtained. The sensing application of such high-Q topological metasurface is also demonstrated, whose figure of merit reaches approximately 145. The proposed strategy and underlying theory can open up new avenues to realize ultrasharp resonances, which can promote numerous potential applications, such as biosensing, optical modulation, and slow-light devices.

Structures of the ß-barrel assembly machine recognizing outer membrane protein substrates.

ß-barrel outer membrane proteins (ß-OMPs) play critical roles in nutrition acquisition, protein import/export, and other fundamental biological processes. The assembly of ß-OMPs in Gram-negative bacteria is mediated by the ß-barrel assembly machinery (BAM) complex, yet its precise mechanism remains elusive. Here, we report two structures of the BAM complex in detergents and in nanodisks, and two crystal structures of the BAM complex with bound substrates. Structural analysis indicates that the membrane compositions surrounding the BAM complex could modulate its overall conformations, indicating low energy barriers between different conformational states and a highly dynamic nature of the BAM complex. Importantly, structures of the BAM complex with bound substrates and the related functional analysis show that the first ß-strand of the BamA ß-barrel (ß1BamA ) in the BAM complex is associated with the last but not the first ß-strand of a ß-OMP substrate via antiparallel ß-strand interactions. These observations are consistent with the ß-signal hypothesis during ß-OMP biogenesis, and suggest that the ß1BamA strand in the BAM complex may interact with the last ß-strand of an incoming ß-OMP substrate upon their release from the chaperone-bound state.

Structure of spinach photosystem II-LHCII supercomplex at 3.2 Å resolution.

During photosynthesis, the plant photosystem II core complex receives excitation energy from the peripheral light-harvesting complex II (LHCII). The pathways along which excitation energy is transferred between them, and their assembly mechanisms, remain to be deciphered through high-resolution structural studies. Here we report the structure of a 1.1-megadalton spinach photosystem II-LHCII supercomplex solved at 3.2 Å resolution through single-particle cryo-electron microscopy. The structure reveals a homodimeric supramolecular system in which each monomer contains 25 protein subunits, 105 chlorophylls, 28 carotenoids and other cofactors. Three extrinsic subunits (PsbO, PsbP and PsbQ), which are essential for optimal oxygen-evolving activity of photosystem II, form a triangular crown that shields the Mn4CaO5-binding domains of CP43 and D1. One major trimeric and two minor monomeric LHCIIs associate with each core-complex monomer, and the antenna-core interactions are reinforced by three small intrinsic subunits (PsbW, PsbH and PsbZ). By analysing the closely connected interfacial chlorophylls, we have obtained detailed insights into the energy-transfer pathways between the antenna and core complexes.

Nanoinfrared Characterization of Bilayer Graphene Conductivity under Dual-Gate Tuning.

Dual-gate tuning on two-dimensional (2D) heterostructures can provide independent control of the carrier concentration and interlayer electrostatic potential, yielding novel electronic and optical properties. In this paper, by utilizing monolayer graphene as both the top gate and a plasmon wavelength magnifier, the optical properties of bilayer graphene (BLG) under dual-gate are quantitatively investigated by nanoinfrared imaging. The hybrid optical modes in the vertically coupled two-layer system are imaged from scattering-type scanning near-field microscopy (s-SNOM). Moreover, plasmon dispersion behaviors under varied dual-gate tuning are explored and explained well with theoretical ones employing tight binding approximation, which reveals the flexibility in individually manipulating the Fermi energy and bandgap. Especially, electron-hole asymmetry in BLG is verified from experiments. Our studies pave route for quantitative near-field investigation of superlattice, topological boundaries, and other emergent phenomena in graphene-based 2D heterostructures.

Effect of charge on protein preferred orientation at the air-water interface in cryo-electron microscopy.

The air-water interface (AWI) tends to adsorb proteins and frequently causes preferred orientation problems in cryo-electron microscopy (cryo-EM). Here, we examined cryo-EM data from protein samples frozen with different detergents and found that both anionic and cationic detergents promoted binding of proteins to the AWI. By contrast, some of the nonionic and zwitterionic detergents tended to prevent proteins from attaching to the AWI. The protein orientation distributions with different anionic detergents were similar and resembled that obtained without detergent. By contrast, cationic detergents gave distinct orientation distributions. Our results indicate that proteins adsorb to charged interface and the negative charge of the AWI plays an important role in adsorbing proteins in the conventional cryo-EM sample preparation. According to these findings, a new method was developed by adding anionic detergent at a concentration between 0.002% and 0.005%. Using this method, the protein particles exhibited a more evenly distributed orientations and still adsorbed to the AWI enabling them embedding in a thin layer of ice with high concentration, which will benefit the cryo-EM structural determination.