A fine-scale Arabidopsis chromatin landscape reveals chromatin conformation-associated transcriptional dynamics.

Plants, as sessile organisms, deploy transcriptional dynamics for adapting to extreme growth conditions such as cold stress. Emerging evidence suggests that chromatin architecture contributes to transcriptional regulation. However, the relationship between chromatin architectural dynamics and transcriptional reprogramming in response to cold stress remains unclear. Here, we apply a chemical-crosslinking assisted proximity capture (CAP-C) method to elucidate the fine-scale chromatin landscape, revealing chromatin interactions within gene bodies closely associated with RNA polymerase II (Pol II) densities across initiation, pausing, and termination sites. We observe dynamic changes in chromatin interactions alongside Pol II activity alterations during cold stress, suggesting local chromatin dynamics may regulate Pol II activity. Notably, cold stress does not affect large-scale chromatin conformations. We further identify a comprehensive promoter-promoter interaction (PPI) network across the genome, potentially facilitating co-regulation of gene expression in response to cold stress. Our study deepens the understanding of chromatin conformation-associated gene regulation in plant response to cold.

Profiling of RNA-binding protein binding sites by in situ reverse transcription-based sequencing.

RNA-binding proteins (RBPs) regulate diverse cellular processes by dynamically interacting with RNA targets. However, effective methods to capture both stable and transient interactions between RBPs and their RNA targets are still lacking, especially when the interaction is dynamic or samples are limited. Here we present an assay of reverse transcription-based RBP binding site sequencing (ARTR-seq), which relies on in situ reverse transcription of RBP-bound RNAs guided by antibodies to identify RBP binding sites. ARTR-seq avoids ultraviolet crosslinking and immunoprecipitation, allowing for efficient and specific identification of RBP binding sites from as few as 20 cells or a tissue section. Taking advantage of rapid formaldehyde fixation, ARTR-seq enables capturing the dynamic RNA binding by RBPs over a short period of time, as demonstrated by the profiling of dynamic RNA binding of G3BP1 during stress granule assembly on a timescale as short as 10 minutes.

A Mechanical Defect Localization and Identification Method for High-Voltage Circuit Breakers Based on the Segmentation of Vibration Signals and Extraction of Chaotic Features.

To address the problem of mechanical defect identification in a high-voltage circuit breaker (HVCB), this paper studies the circuit breaker vibration signal and proposes a method of feature extraction based on phase-space reconstruction of the vibration substages. To locate mechanical defects in circuit breakers, vibration signals are divided into different substages according to the time sequence of the parts of the circuit breakers. The largest Lyapunov exponent (LLE) of the vibration signals' substages is calculated, and then the substages are reconstructed in high-dimensional phase space. The geometric features of the phase trajectory mean center distance (MCD) and vector diameter offset (VDO) are calculated, and the LLE, MCD, and VDO are selected as the three fault identification features of the vibration substages. The eigenvalue anomaly rate of each substage of the vibration signal under defect state are calculated and analyzed to locate the vibration substage of the mechanical defect. Finally, a fault diagnosis model is constructed by a support vector machine (SVM), and the common mechanical defects of circuit breakers simulated in the laboratory are effectively identified.

Research on Circuit Breaker Operating Mechanism Fault Diagnosis Method Combining Global-Local Feature Extraction and KELM.

In response to the lack of generality in feature extraction using modal decomposition methods and the susceptibility of diagnostic performance to parameter selection in traditional mechanical fault diagnosis of high-voltage circuit breaker operating mechanisms, this paper proposes a Global-Local feature extraction method based on Generalized S-Transform (S-Translate) combined with Gray Level Co-Occurrence Matrix (GLCM) and complemented by Maximum Relevance and Minimum Redundancy (mRMR) feature selection. The GL (Global-Local)-mRMR-KELM fault diagnosis model is proposed, which employs the Kernel Extreme Learning Machine (KELM). In this model, the original time-frequency domain features and the time-frequency features of the Generalized S-Transform matrix of vibration signals under different states of the circuit breaker are first extracted as global features. Then, the GLCM is obtained to extract texture features as local features. Finally, the mRMR and KELM are comprehensively applied to perform feature selection and classification on the dataset, thereby accomplishing the fault diagnosis of the circuit breaker's operating mechanism. In this study, the 72.5 kV SF6 circuit breaker operating mechanism is taken as the research object, and three types of mechanical faults are simulated to obtain a vibration signal. Experimental results verify the effectiveness of the proposed GL-mRMR-KELM model, achieving a diagnostic accuracy of 96%. This research provides a feasible approach for the fault diagnosis of circuit breaker operating mechanisms.

FTO mediates LINE1 m6A demethylation and chromatin regulation in mESCs and mouse development.

N6-methyladenosine (m6A) is the most abundant internal modification on mammalian messenger RNA. It is installed by a writer complex and can be reversed by erasers such as the fat mass and obesity-associated protein FTO. Despite extensive research, the primary physiological substrates of FTO in mammalian tissues and development remain elusive. Here, we show that FTO mediates m6A demethylation of long-interspersed element-1 (LINE1) RNA in mouse embryonic stem cells (mESCs), regulating LINE1 RNA abundance and the local chromatin state, which in turn modulates the transcription of LINE1-containing genes. FTO-mediated LINE1 RNA m6A demethylation also plays regulatory roles in shaping chromatin state and gene expression during mouse oocyte and embryonic development. Our results suggest broad effects of LINE1 RNA m6A demethylation by FTO in mammals.

Nonsegmented Negative-Sense RNA Viruses Utilize N6-Methyladenosine (m6A) as a Common Strategy To Evade Host Innate Immunity.

N6-Methyladenosine (m6A) is the most abundant internal RNA modification catalyzed by host RNA methyltransferases. As obligate intracellular parasites, many viruses acquire m6A methylation in their RNAs. However, the biological functions of viral m6A methylation are poorly understood. Here, we found that viral m6A methylation serves as a molecular marker for host innate immunity to discriminate self from nonself RNA and that this novel biological function of viral m6A methylation is universally conserved in several families in nonsegmented negative-sense (NNS) RNA viruses. Using m6A methyltransferase (METTL3) knockout cells, we produced m6A-deficient virion RNAs from the representative members of the families Pneumoviridae, Paramyxoviridae, and Rhabdoviridae and found that these m6A-deficient viral RNAs triggered significantly higher levels of type I interferon compared to the m6A-sufficient viral RNAs, in a RIG-I-dependent manner. Reconstitution of the RIG-I pathway revealed that m6A-deficient virion RNA induced higher expression of RIG-I, bound to RIG-I more efficiently, enhanced RIG-I ubiquitination, and facilitated RIG-I conformational rearrangement and oligomerization. Furthermore, the m6A binding protein YTHDF2 is essential for suppression of the type I interferon signaling pathway, including by virion RNA. Collectively, our results suggest that several families in NNS RNA viruses acquire m6A in viral RNA as a common strategy to evade host innate immunity.IMPORTANCE The nonsegmented negative-sense (NNS) RNA viruses share many common replication and gene expression strategies. There are no vaccines or antiviral drugs for many of these viruses. We found that representative members of the families Pneumoviridae, Paramyxoviridae, and Rhabdoviridae among the NNS RNA viruses acquire m6A methylation in their genome and antigenome as a means to escape recognition by host innate immunity via a RIG-I-dependent signaling pathway. Viral RNA lacking m6A methylation induces a significantly higher type I interferon response than m6A-sufficient viral RNA. In addition to uncovering m6A methylation as a common mechanism for many NNS RNA viruses to evade host innate immunity, this study discovered a novel strategy to enhance type I interferon responses, which may have important applications in vaccine development, as robust innate immunity will likely promote the subsequent adaptive immunity.

Development of a C2-Symmetric Chiral aza Spirocyclic Diol.

A C2-symmetric chiral spirocyclic diol aza-SPINOL containing a spirooxindole scaffold has been designed, synthesized, and optically resolved. The product could be synthesized on gram scale in an overall yield of 22%. Moreover, elaborations of aza-SPINOL to other chiral ligands as well as the preliminary investigation of the related bisphosphine ligand in the desymmetrization of bisallylic amide were reported.

Ruthenium(II)-Catalyzed Asymmetric Inert C-H Bond Activation Assisted by a Chiral Transient Directing Group.

A ruthenium(II)-catalyzed asymmetric intramolecular hydroarylation assisted by a chiral transient directing group has been developed. A series of 2,3-dihydrobenzofurans bearing chiral all-carbon quaternary stereocenters have been prepared in remarkably high yields (up to 98 %) and enantioselectivities (up to >99 % ee). By this methodology, a novel asymmetric total synthesis of CB2 receptor agonist MDA7 has been successfully developed.