Henipaviruses and lyssaviruses target nucleolar treacle protein and regulate ribosomal RNA synthesis.

The nucleolus is a common target of viruses and viral proteins, but for many viruses the functional outcomes and significance of this targeting remains unresolved. Recently, the first intranucleolar function of a protein of a cytoplasmically-replicating negative-sense RNA virus (NSV) was identified, with the finding that the matrix (M) protein of Hendra virus (HeV) (genus Henipavirus, family Paramyxoviridae) interacts with Treacle protein within nucleolar subcompartments and mimics a cellular mechanism of the nucleolar DNA-damage response (DDR) to suppress ribosomal RNA (rRNA) synthesis. Whether other viruses utilise this mechanism has not been examined. We report that sub-nucleolar Treacle targeting and modulation is conserved between M proteins of multiple Henipaviruses, including Nipah virus and other potentially zoonotic viruses. Furthermore, this function is also evident for P3 protein of rabies virus, the prototype virus of a different RNA virus family (Rhabdoviridae), with Treacle depletion in cells also found to impact virus production. These data indicate that unrelated proteins of viruses from different families have independently developed nucleolar/Treacle targeting function, but that modulation of Treacle has distinct effects on infection. Thus, subversion of Treacle may be an important process in infection by diverse NSVs, and so could provide novel targets for antiviral approaches with broad specificity.

Gas-Responsive and Self-Powered Visual Composite Langmuir-Blodgett Films for Ultrathin Gas Sensors.

The complex and variable environments are challenging the development of related detection and analysis. Ammonia (NH3) and hydrogen chloride (HCl) gases are both commonly used in industry, but they are considered to be toxic and corrosive substances that can threaten human health and the environment. Therefore, it is necessary here to develop a convenient, sensitive, and reliable sensor device for acid-alkali gas detection. Herein, we propose the synthesis strategy of an ultrathin film gas sensor based on the pH-responsive, self-powered, and visible composite Langmuir-Blodgett (LB) films. In our work, the LB films with nanometric thicknesses are obtained based on the sensitive materials of two novel carbazole structural sensitizers (abbreviated as CS-35 and CS-37) and several dye molecules. The composite LB films are formed with Carbazole samples and dye molecules through hydrogen bonding, π-π stacking, synergistic electrostatic interactions, and hydrophobic interactions, existing as J-aggregate or H-aggregate. The formation of high-quality and uniform Langmuir films is confirmed with transmission electron microscope (TEM), UV-vis spectrum, atomic force microscopy (AFM), and other measurements. In addition, based on the simple protonation and deprotonation, the prepared LB films can be assembled into a visual sensor for the response of pH gases. The response is confirmed by the study of ultraviolet spectroscopy and electrical output in vertical contact separation mode, which potentially unlocks a sustainable future for the application of ultrathin self-powered gas sensors.

FKBP8 interact with classical swine fever virus NS5A protein and promote virus RNA replication.

The non-structural 5A (NS5A) protein of classical swine fever virus (CSFV) is proven to be involved in viral replication and can also modulate cellular signaling and host cellular responses via to its ability to interact with various cellular proteins. FKBP8 is also reported to promote virus replication. Here, we show that NS5A specifically interacts with FKBP8 through coimmunoprecipitation and GST-pulldown studies. Additionally, confocal microscopy study showed that NS5A and FKBP8 colocalized in the cytoplasm. Overexpression of FKBP8 via the eukaryotic expression plasmid pDsRED N1 significantly promoted viral RNA synthesis. The cells knockdown of FKBP8 by lentivirus-mediated shRNA markedly decreased the virus replication when infected with CSFV. These data suggest that FKBP8 plays a critical role in the viral life cycle, particularly during the virus RNA replication period. The investigation of FKBP8 protein functions may be beneficial for developing new strategies to treat CSFV infection.

Feed-forward stimulation of CAMK2 by the oncogenic pseudokinase PEAK1 generates a therapeutically 'actionable' signalling axis in triple negative breast cancer.

The PEAK family of pseudokinases, comprising PEAK1-3, are signalling scaffolds that play oncogenic roles in several poor prognosis human cancers, including triple negative breast cancer (TNBC). However, therapeutic targeting of pseudokinases is challenging due to their lack of catalytic activity. To address this, we screened for PEAK1 effectors by affinity purification and mass spectrometry, identifying calcium/calmodulin-dependent protein kinase 2 (CAMK2)D and CAMK2G. PEAK1 promoted CAMK2D/G activation in TNBC cells via a novel feed-forward mechanism involving PEAK1/PLCγ1/Ca 2+ signalling and direct binding via a consensus CAMK2 interaction motif in the PEAK1 N-terminus. In turn, CAMK2 phosphorylated PEAK1 to enhance association with PEAK2, which is critical for PEAK1 oncogenic signalling. To achieve pharmacologic targeting of PEAK1/CAMK2, we repurposed RA306, a second generation CAMK2 inhibitor under pre-clinical development for treatment of cardiovascular disease. RA306 demonstrated on-target activity against CAMK2 in TNBC cells and inhibited PEAK1-enhanced migration and invasion in vitro . Moreover, RA306 significantly attenuated TNBC xenograft growth and blocked metastasis in a manner mirrored by CRISPR-mediated PEAK1 ablation. Overall, these studies establish PEAK1 as a critical cell signalling nexus, identify a novel mechanism for regulation of Ca 2+ signalling and its integration with tyrosine kinase signals, and identify CAMK2 as a therapeutically 'actionable' target downstream of PEAK1.

Fermentation Quality, In Vitro Digestibility, and Aerobic Stability of Ensiling Spent Mushroom Substrate with Microbial Additives.

This experiment investigated the effects of lactic acid bacteria and cellulase on the fermentation quality, in vitro digestibility, and aerobic stability of Flammulina velutipes spent mushroom substrate silage (F-silage) and Pleurotus eryngii spent mushroom substrate silage (P-silage). Silage treatments included groups without any additives (control), with lactic acid bacteria (L), with cellulase (E), and with lactic acid bacteria and cellulase (M). Data analysis was performed using independent sample t-test and analysis of variance. After 45 days of ensiling, the pH in F-silage and P-silage from the L, E, and M groups were lower than those in the control group (p < 0.05). The pH, acetic acid (AA), and propionic acid (PA) levels in P-silage were lower than those in F-silage, and the LA content in P-silage was higher than that in F-silage (p < 0.05). Compared with the control, the E treatment increased in vitro neutral detergent fibre digestibility (IVNDFD) and in vitro acid detergent fibre digestibility (IVADFD) in F-silage and P-silage (p < 0.05). The aerobic stability of F-silage inoculated with L increased (p < 0.05) by 24 h compared to the control. The aerobic stability of P-silage inoculated with M increased (p < 0.05) by 6 h compared to the control. The improvement in fermentation quality and aerobic stability is extremely large in terms of applying M in F-silage and P-silage. The E is effective in improving the in vitro digestibility of P-silage. The research results provide a theoretical basis for the production of high-quality spent mushroom substrate fermented feed.

Viral regulation of host cell biology by hijacking of the nucleolar DNA-damage response.

Recent studies indicate that nucleoli play critical roles in the DNA-damage response (DDR) via interaction of DDR machinery including NBS1 with nucleolar Treacle protein, a key mediator of ribosomal RNA (rRNA) transcription and processing. Here, using proteomics, confocal and single molecule super-resolution imaging, and infection under biosafety level-4 containment, we show that this nucleolar DDR pathway is targeted by infectious pathogens. We find that the matrix proteins of Hendra virus and Nipah virus, highly pathogenic viruses of the Henipavirus genus in the order Mononegavirales, interact with Treacle and inhibit its function, thereby silencing rRNA biogenesis, consistent with mimicking NBS1-Treacle interaction during a DDR. Furthermore, inhibition of Treacle expression/function enhances henipavirus production. These data identify a mechanism for viral modulation of host cells by appropriating the nucleolar DDR and represent, to our knowledge, the first direct intranucleolar function for proteins of any mononegavirus.

Synthesis, characterization, and field-effect transistor properties of carbazolenevinylene oligomers: from linear to cyclic architectures.

Two cyclic carbazolenevinylene dimers 1 and 2 were synthesized by McMurry coupling reactions. A linear compound 3 was also prepared for comparison. Compounds 1-3 were fully characterized by means of NMR spectroscopy, HRMS, elemental analysis, and UV/Vis absorption spectroscopy. Quantum chemical simulations showed that the cyclic compounds possessed smaller HOMO-LUMO gaps and more extended conjugation. The UV/Vis absorption spectra of the cyclic compounds showed blueshifts compared with that of the linear compound 3. Time-dependent DFT (TD-DFT) analysis revealed that this was due to the different selection rules for molecules with cyclic and linear architectures. The cyclic conformation also significantly affected the molecular ordering in the solid state. The X-ray crystal structure of 1 showed partial pi-pi overlapping between the adjacent molecules. Thin films of 1-3 were fabricated by the vacuum-deposition method on Si/SiO(2) substrates. Multicrystalline thin films were obtained from compounds 1 and 2, but only amorphous thin films could be obtained for the linear compound 3. Another important difference between the cyclic and linear compounds was the reduced reorganization energy for the cyclic compounds. These two facts have resulted in improved field-effect transistor (FET) mobilities for the cyclic compounds compared with the linear compound. In addition, as the substrate temperature has a significant influence on the morphology and the degree of crystallinity of the thin films deposited, the device performance could be optimized by varying the substrate temperature. The FET devices based on 2 gave the highest mobility of 0.013 cm(2) V(-1) s(-1). The results showed that carbazole derivatives with cyclic structures might make better FETs.

Novel diethynylcarbazole macrocycles: synthesis and optoelectronic properties.

Diethynylcarbazole macrocycles 1b and 2b have been synthesized by oxidative coupling of appropriate precursors. In particular, macrocycle 2b was prepared by bimolecular Pd-catalyzed oxidative coupling in 35% isolated yield. The spectroscopic properties of these macrocycles and their precursors were measured in detail. The films of these macrocycles by the dipping method and the Langmuir-Blodgett technique were fabricated to study their photoinduced charge-transfer properties. A rapid and steady cathodic photocurrent of these films was produced in a three-electrode cell when irradiated with white light. A possible mechanism of the photoinduced electron-transfer pathway was suggested.