mTORC1 activity is supported by spatial association with focal adhesions

The mammalian target of rapamycin complex 1 (mTORC1) integrates mitogenic and stress signals to control growth and metabolism. Activation of mTORC1 by amino acids and growth factors involves recruitment of the complex to the lysosomal membrane and is further supported by lysosome distribution to the cell periphery. Here, we show that translocation of lysosomes toward the cell periphery brings mTORC1 into proximity with focal adhesions (FAs). We demonstrate that FAs constitute discrete plasma membrane hubs mediating growth factor signaling and amino acid input into the cell. FAs, as well as the translocation of lysosome-bound mTORC1 to their vicinity, contribute to both peripheral and intracellular mTORC1 activity. Conversely, lysosomal distribution to the cell periphery is dispensable for the activation of mTORC1 constitutively targeted to FAs. This study advances our understanding of spatial mTORC1 regulation by demonstrating that the localization of mTORC1 to FAs is both necessary and sufficient for its activation by growth-promoting stimuli.     

Sterol and oxysterol synthases near the ciliary base activate the Hedgehog pathway

Vertebrate Hedgehog signals are transduced through the primary cilium, a specialized lipid microdomain that is required for Smoothened activation. Cilia-associated sterol and oxysterol lipids bind to Smoothened to activate the Hedgehog pathway, but how ciliary lipids are regulated is incompletely understood. Here we identified DHCR7, an enzyme that produces cholesterol, activates the Hedgehog pathway, and localizes near the ciliary base. We found that Hedgehog stimulation negatively regulates DHCR7 activity and removes DHCR7 from the ciliary microenvironment, suggesting that DHCR7 primes cilia for Hedgehog pathway activation. In contrast, we found that Hedgehog stimulation positively regulates the oxysterol synthase CYP7A1, which accumulates near the ciliary base and produces oxysterols that promote Hedgehog signaling in response to pathway activation. Our results reveal that enzymes involved in lipid biosynthesis in the ciliary microenvironment promote Hedgehog signaling, shedding light on how ciliary lipids are established and regulated to transduce Hedgehog signals.     

水痘-带状疱疹病毒ORF43蛋白的单克隆抗体制备及初步研究

水痘-带状疱疹病毒(Varicella-zoster virus,VZV)是引起水痘和带状疱疹这两种临床表现不同病症的共同致病原,其基因组中ORF43是VZV在宿主细胞中复制的必需基因,但目前尚无针对VZV ORF43编码蛋白性质与功能的研究报道.本研究目的 是制备抗VZV ORF43单克隆抗体,以初步研究该蛋白在细胞内的表达与分布情况.本研究构建了VZV ORF43蛋白的原核表达质粒并在大肠杆菌中进行了该蛋白的表达,纯化蛋白免疫小鼠后,使用杂交瘤技术及克隆化筛选,获得一株特异性强、反应性好的抗VZV ORF43单克隆抗体2D3.本研究使用该抗体鉴定出VZV ORF43蛋白在质粒转染细胞与病毒感染细胞中表达的分子量大小均约为70kD,但分别呈现出细胞质和细胞核的不同亚细胞定位.以上工作为后续进一步探究该蛋白在VZV感染与致病过程中的作用奠定了基础.     

模拟干旱胁迫处理下北柴胡种苗MYC2基因的调控研究

该研究以北柴胡一年生种苗为材料,使用质量分数为10%、20%的PEG6000营养液进行模拟干旱胁迫实验,检测北柴胡根内源信号物质OPR、JA含量,转录因子BcMYC2和柴胡皂苷生物合成途径中4个关键酶基因HMGR、IPPI、FPS、β AS表达量,以及柴胡皂苷a、d含量,探究模拟干旱胁迫下茉莉酸信号通路调控BcMYC2进而影响柴胡皂苷生物合成的机制。结果表明：(1) PEG6000模拟干旱胁迫处理北柴胡种苗后,20% PEG6000处理组根内OPR含量在2 h时出现峰值,10% PEG6000处理组的根内OPR含量在6 h时出现峰值;两个胁迫组内源JA含量均在2 h时出现峰值。(2)两个胁迫处理组北柴胡根内BcMYC2相对表达量均在2 h处出现峰值,之后的2~4 h时间段内大幅度下降,且20% PEG6000处理组的BcMYC2相对表达量高于10% PEG6000处理组;其余4个柴胡皂苷生物合成途径关键酶基因HMGR、IPPI、FPS、β AS的相对表达量均在4 h时出现峰值,晚于BcMYC2相对表达量出现峰值的时间。(3)经模拟干旱胁迫处理后,北柴胡根内的柴胡皂苷含量在36 d内逐渐上升;在处理36 d时,20% PEG6000处理组的柴胡皂苷含量略高于10% PEG6000处理组,且两处理组均显著高于对照组。研究发现,经PEG6000模拟干旱处理后,北柴胡根内信号物质OPR及JA含量提高,促进BcMYC2的表达,进而提高柴胡皂苷生物合成途径中的关键酶基因的表达,最终显著提高了根内柴胡皂苷的含量。     

Sustainability of SARS-CoV-2 Induced Humoral Immune Responses in COVID-19 Patients from Hospitalization to Convalescence Over Six Months

Virologica Sinica - Understanding the persistence of antibody in convalescent COVID-19 patients may help to answer the current major concerns such as the risk of reinfection, the protection period...     

The Efficacy of a Live Attenuated TW I-Type Infectious Bronchitis Virus Vaccine Candidate

Virologica Sinica - Infectious bronchitis (IB) is a highly contagious avian disease caused by infection with infectious bronchitis virus (IBV), which seriously affects the development of the global...     

A practical guide to amplicon and metagenomic analysis of microbiome data

Advances in high-throughput sequencing(HTS)have fostered rapid developments in the field of microbiome research,and massive microbiome datasets are now being generated.However,the diversity of software tools and the complexity of analysis pipelines make it difficult to access this field.Here,we systematically summarize the advantages and limitations of micro-biome methods.Then,we recommend specific pipelines for amplicon and metagenomic analyses,and describe commonly-used software and databases,to help researchers select the appropriate tools.Furthermore,we introduce statistical and visualization methods suit-able for microbiome analysis,including alpha-and beta-diversity,taxonomic composition,difference compar-isons,correlation,networks,machine learning,evolu-tion,source tracing,and common visualization styles to help researchers make informed choices.Finally,a step-by-step reproducible analysis guide is introduced.We hope this review will allow researchers to carry out data analysis more effectively and to quickly select the appropriate tools in order to efficiently mine the bio-logical significance behind the data.     

Is oral microbiome of children able to maintain resistance and functional stability in response to short-term interference of ingesta?

Dear Editor, A series of studies had focused on the ecological stability of human microbiome (Lozupone et al.,2012;Faith et al.,2013;Moya and Ferrer,2016).Despite the continuous perturbation and the highly personalized composition within the human microbiome (Human Microbiome Project,2012),healthy adults stably maintain their microbial communities in terms of space and time (Faith et al.,2013;Moya and Ferrer,2016;Oh et al.,2016).This stability is proved to be critical for the well-being of human body (Lozupone et al.,2012).On the contrary,major shifts in microbial community composition are often related to diseases (Lynch and Pedersen,2016).     

Inhibition of cyclooxygenase-2 enhanced intestinal epithelial homeostasis via suppressing β-catenin signalling pathway in experimental liver fibrosis

The intestinal barrier dysfunction is crucial for the development of liver fibrosis but can be disturbed by intestinal chronic inflammation characterized with cyclooxygenase-2 (COX-2) expression. This study focused on the unknown mechanism by which COX-2 regulates intestinal epithelial homeostasis in liver fibrosis. The animal models of liver fibrosis induced with TAA were established in rats and in intestinal epithelial–specific COX-2 knockout mice. The impacts of COX-2 on intestinal epithelial homeostasis via suppressing β-catenin signalling pathway were verified pharmacologically and genetically in vivo. A similar assumption was tested in Ls174T cells with goblet cell phenotype in vitro. Firstly, disruption of intestinal epithelial homeostasis in cirrhotic rats was ameliorated by celecoxib, a selective COX-2 inhibitor. Then, β-catenin signalling pathway in cirrhotic rats was associated with the activation of COX-2. Furthermore, intestinal epithelial–specific COX-2 knockout could suppress β-catenin signalling pathway and restore the disruption of ileal epithelial homeostasis in cirrhotic mice. Moreover, the effect of COX-2/PGE2 was dependent on the β-catenin signalling pathway in Ls174T cells. Therefore, inhibition of COX-2 may enhance intestinal epithelial homeostasis via suppression of the β-catenin signalling pathway in liver fibrosis.