Video_1_Spatio-temporal spread and evolution of influenza A (H7N9) viruses.MP4

The influenza A (H7N9) virus has been seriously concerned for its potential to cause an influenza pandemic. To understand the spread and evolution process of the virus, a spatial and temporal Bayesian evolutionary analysis was conducted on 2,052 H7N9 viruses isolated during 2013 and 2018. It revealed that the H7N9 virus was probably emerged in a border area of Anhui Province in August 2012, approximately 6 months earlier than the first human case reported. Two major epicenters had been developed in the Yangtze River Delta and Peral River Delta regions by the end of 2013, and from where the viruses have also spread to other regions at an average speed of 6.57 km/d. At least 24 genotypes showing have been developed and each of them showed a distinct spatio-temporal distribution pattern. Furthermore, A random forest algorithm-based model has been developed to predict the occurrence risk of H7N9 virus. The model has a high overall forecasting precision (> 97%) and the monthly H7N9 occurrence risk for each county of China was predicted. These findings provide new insights for a comprehensive understanding of the origin, evolution, and occurrence risk of H7N9 virus. Moreover, our study also lays a theoretical basis for conducting risk-based surveillance and prevention of the disease.</p

Video_3_Spatio-temporal spread and evolution of influenza A (H7N9) viruses.MP4

The influenza A (H7N9) virus has been seriously concerned for its potential to cause an influenza pandemic. To understand the spread and evolution process of the virus, a spatial and temporal Bayesian evolutionary analysis was conducted on 2,052 H7N9 viruses isolated during 2013 and 2018. It revealed that the H7N9 virus was probably emerged in a border area of Anhui Province in August 2012, approximately 6 months earlier than the first human case reported. Two major epicenters had been developed in the Yangtze River Delta and Peral River Delta regions by the end of 2013, and from where the viruses have also spread to other regions at an average speed of 6.57 km/d. At least 24 genotypes showing have been developed and each of them showed a distinct spatio-temporal distribution pattern. Furthermore, A random forest algorithm-based model has been developed to predict the occurrence risk of H7N9 virus. The model has a high overall forecasting precision (> 97%) and the monthly H7N9 occurrence risk for each county of China was predicted. These findings provide new insights for a comprehensive understanding of the origin, evolution, and occurrence risk of H7N9 virus. Moreover, our study also lays a theoretical basis for conducting risk-based surveillance and prevention of the disease.</p

Video_2_Spatio-temporal spread and evolution of influenza A (H7N9) viruses.MP4

The influenza A (H7N9) virus has been seriously concerned for its potential to cause an influenza pandemic. To understand the spread and evolution process of the virus, a spatial and temporal Bayesian evolutionary analysis was conducted on 2,052 H7N9 viruses isolated during 2013 and 2018. It revealed that the H7N9 virus was probably emerged in a border area of Anhui Province in August 2012, approximately 6 months earlier than the first human case reported. Two major epicenters had been developed in the Yangtze River Delta and Peral River Delta regions by the end of 2013, and from where the viruses have also spread to other regions at an average speed of 6.57 km/d. At least 24 genotypes showing have been developed and each of them showed a distinct spatio-temporal distribution pattern. Furthermore, A random forest algorithm-based model has been developed to predict the occurrence risk of H7N9 virus. The model has a high overall forecasting precision (> 97%) and the monthly H7N9 occurrence risk for each county of China was predicted. These findings provide new insights for a comprehensive understanding of the origin, evolution, and occurrence risk of H7N9 virus. Moreover, our study also lays a theoretical basis for conducting risk-based surveillance and prevention of the disease.</p

MOESM1 of EZH2 promotes DNA replication by stabilizing interaction of POLδ and PCNA via methylation-mediated PCNA trimerization

Additional file 1: Figure S1 (related to Fig. 1). A: Immunoblot shows the H3K27me3 level in EZH2-knockdown hDPCs. B: The morphology of hDPCs with EZH2 knockdown. C: Quantification of colony-forming ability of hDPCs with EZH2 knockdown. D: Charts represent genes in the cell cycle RNA-sequencing assay that are involved in DNA replication based on the AmiGO 2 database. (ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001). Scale bar represents 100 μ

MOESM3 of EZH2 promotes DNA replication by stabilizing interaction of POLδ and PCNA via methylation-mediated PCNA trimerization

Additional file 3: Figure S3. Uncut immunoblots from the immunoprecipitation experiment

MOESM2 of EZH2 promotes DNA replication by stabilizing interaction of POLδ and PCNA via methylation-mediated PCNA trimerization

Additional file 2: Figure S2 (related to Fig. 2). A: Flow cytometry shows the cell cycle distribution of hDPCs from one donor at the indicated time points after release from serum deprivation. B: Immunoblot demonstrates the expression of CYCLIN D1 and CYCLIN A2, indicating G1 phase and S phase, respectively. α-TUBULIN was used a loading control of whole lysate. C: Statistical analysis of the EZH2-positive ratio (Left, n = 3) and Pearson’s correlation coefficient of EZH2 with DAPI (right, each dot represents one cell). D: Statistical analysis of the PLA-positive ratio (left, n = 5). The PLA signals are associated with DAPI on a single-cell basis (***P < 0.001

Data_Sheet_1_The W-Acidic Motif of Histidine Kinase WalK Is Required for Signaling and Transcriptional Regulation in Streptococcus mutans.docx

In Streptococcus mutans, we find that the histidine kinase WalK possesses the longest C-terminal tail (CTT) among all 14 TCSs, and this tail plays a key role in the interaction of WalK with its response regulator WalR. We demonstrate that the intrinsically disordered CTT is characterized by a conserved tryptophan residue surrounded by acidic amino acids. Mutation in the tryptophan not only disrupts the stable interaction, but also impairs the efficient phosphotransferase and phosphatase activities of WalRK. In addition, the tryptophan is important for WalK to compete with DNA containing a WalR binding motif for the WalR interaction. We further show that the tryptophan is important for in vivo transcriptional regulation and bacterial biofilm formation by S. mutans. Moreover, Staphylococcus aureus WalK also has a characteristic CTT, albeit relatively shorter, with a conserved W-acidic motif, that is required for the WalRK interaction in vitro. Together, these data reveal that the W-acidic motif of WalK is indispensable for its interaction with WalR, thereby playing a key role in the WalRK-dependent signal transduction, transcriptional regulation and biofilm formation.</p

Mechanistic Insights Revealed by the Crystal Structure of a Histidine Kinase with Signal Transducer and Sensor Domains

<div><p>Two-component systems (TCSs) are important for the adaptation and survival of bacteria and fungi under stress conditions. A TCS is often composed of a membrane-bound sensor histidine kinase (SK) and a response regulator (RR), which are relayed through sequential phosphorylation steps. However, the mechanism for how an SK is switched on in response to environmental stimuli remains obscure. Here, we report the crystal structure of a complete cytoplasmic portion of an SK, VicK from <i>Streptococcus mutans</i>. The overall structure of VicK is a long-rod dimer that anchors four connected domains: HAMP, Per-ARNT-SIM (PAS), DHp, and catalytic and ATP binding domain (CA). The HAMP, a signal transducer, and the PAS domain, major sensor, adopt canonical folds with dyad symmetry. In contrast, the dimer of the DHp and CA domains is asymmetric because of different helical bends in the DHp domain and spatial positions of the CA domains. Moreover, a conserved proline, which is adjacent to the phosphoryl acceptor histidine, contributes to helical bending, which is essential for the autokinase and phosphatase activities. Together, the elegant architecture of VicK with a signal transducer and sensor domain suggests a model where DHp helical bending and a CA swing movement are likely coordinated for autokinase activation.</p> </div

Analyses for autokinase activation.

<p>(A) Structural alignment of two VicK monomers in reference to the DHp domain. The alignment was carried out as described in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001493#pbio-1001493-g004" target="_blank">Figure 4A</a>. The vertical movement is indicated with a black arrow in parallel to the DHp domain while a grey arrow indicates a rotation. (B) The VicK CA domain (magenta) aligned to <i>B. subtilis</i> YycG (light blue) in the presence of ATP (green and gold sticks). His217 is represented by green sticks and several other residues are shown in magenta and gold sticks. The contacts between the distances of 2.5–3.5 Å are highlighted with yellow dashed lines and those between 3.5–4.5 Å with grey dashed lines. (C) Residues critical for the interaction between the DHp and active CA domains. The VicK Cα chain is colored in gray ribbon and residues from the CA domain critical for this interface in cyan sticks. The critical residues from the DHp domain are highlighted in grey sticks. The right panel is a flip-over view of the left. (D) Mutations of key residues in the active interface affected autokinase activity. The amount of wt VicK and mutants used in each reaction were analyzed by 15% SDS-PAGE and stained by coomassie blue as shown in the bottom gel. A protein marker served as a background control (lane 6). The gel containing phosphorylation forms of VicK was exposed to an X-ray film as shown in the top gel.</p

Proline and threonine are essential for VicK phosphatase activity.

<p>(A) VicR dephosphorylation examined in PMS. Phosphorylated VicR was treated with the same amount of VicK wt or its mutants as shown in the bottom gel. Native and phosphorylated VicR were separated in PMS as labeled on the right of the top two gels, where in one series of reactions additional 5 mM ATP was used (the top 1 gel). (B) VicK phosphatase activity analyzed by HPLC. AcP-treated VicR was incubated with VicK in the reaction buffer with 5 mM ATP (right) and without ATP (left). VicK phosphatase activity was analyzed by the phosphorylation state of VicR on HPLC.</p