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目的 研究严重急性呼吸综合征冠状病毒2(SARS-CoV-2)膜蛋白对宿主细胞mRNA前体(pre-mRNA)3"非翻译区(UTR)加工的影响。方法 本研究以人肺上皮细胞系A549为模型,利用瞬时转染在细胞内过表达SARS-CoV-2膜蛋白;利用RNA-Seq测序技术及生物信息学分析方法,系统性描绘宿主细胞选择性多聚腺苷酸化(alternative polyadenylation,APA)事件;Metascape数据库对发生显著APA变化的基因进行功能富集分析;RT-qPCR验证靶基因3"UTR长度变化;蛋白质免疫印迹(Western blot)检测目的蛋白表达水平。结果 SARS-CoV-2膜蛋白外源表达后宿主细胞内共813个基因发生显著APA变化。GO和KEGG分析显示,差异APA基因广泛参与有丝分裂细胞周期、调节细胞应激等生物过程,涉及病毒感染和蛋白质加工等。从中进一步筛选出AKT1基因,在IGV软件中显示3"UTR延长;RT-qPCR验证AKT1基因的3"UTR长度变化趋势;Western blot结果显示AKT1蛋白磷酸化水平增加。结论 SARS-CoV-2膜蛋白潜在影响宿主pre-mRNA的3"UTR加工,其中参与多种病毒性生物过程的AKT1基因 3"UTR延长,且其编码的蛋白质功能在细胞内被激活。 相似文献
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Peng Wang Ronghua Luo Min Zhang Yaqing Wang Tianzhang Song Tingting Tao Zhongyu Li Lin Jin Hongyi Zheng Wenwen Chen Mengqian Zhao Yongtang Zheng Jianhua Qin 《Cell death & disease》2020,11(12)
COVID-19, caused by SARS-CoV-2, is an acute and rapidly developing pandemic, which leads to a global health crisis. SARS-CoV-2 primarily attacks human alveoli and causes severe lung infection and damage. To better understand the molecular basis of this disease, we sought to characterize the responses of alveolar epithelium and its adjacent microvascular endothelium to viral infection under a co-culture system. SARS-CoV-2 infection caused massive virus replication and dramatic organelles remodeling in alveolar epithelial cells, alone. While, viral infection affected endothelial cells in an indirect manner, which was mediated by infected alveolar epithelium. Proteomics analysis and TEM examinations showed viral infection caused global proteomic modulations and marked ultrastructural changes in both epithelial cells and endothelial cells under the co-culture system. In particular, viral infection elicited global protein changes and structural reorganizations across many sub-cellular compartments in epithelial cells. Among the affected organelles, mitochondrion seems to be a primary target organelle. Besides, according to EM and proteomic results, we identified Daurisoline, a potent autophagy inhibitor, could inhibit virus replication effectively in host cells. Collectively, our study revealed an unrecognized cross-talk between epithelium and endothelium, which contributed to alveolar–capillary injury during SARS-CoV-2 infection. These new findings will expand our understanding of COVID-19 and may also be helpful for targeted drug development.Subject terms: Mechanisms of disease, Viral infection 相似文献
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Integrated species delimitation and conservation implications of an endangered weevil Pachyrhynchus sonani (Coleoptera: Curculionidae) in Green and Orchid Islands of Taiwan 下载免费PDF全文
YEN‐TING CHEN HUI‐YUN TSENG MING‐LUEN JENG YONG‐CHAO SU WEN‐SAN HUANG CHUNG‐PING LIN 《Systematic Entomology》2017,42(4):796-813
Oceanic islands are productive habitats for generating new species and high endemism, which is primarily due to their geographical isolation, smaller population sizes and local adaptation. However, the short divergence times and subtle morphological or ecological divergence of insular organisms may obscure species identity, so the cryptic endemism on islands may be underestimated. The endangered weevil Pachyrhynchus sonani Kôno (Coleoptera: Curculionidae: Entiminae: Pachyrhynchini) is endemic to Green Island and Orchid Island of the Taiwan‐Luzon Archipelago and displays widespread variation in coloration and host range, thus raising questions regarding its species boundaries and degree of cryptic diversity. We tested the species boundaries of P. sonani using an integrated approach that combined morphological (body size and shape, genital shape, coloration and cuticular scale), genetic (four genes and restriction site‐associated DNA sequencing, RAD‐seq) and ecological (host range and distribution) diversity. The results indicated that all the morphological datasets for male P. sonani, except for the colour spectrum, reveal overlapping but statistically significant differences between islands. In contrast, the morphology of the female P. sonani showed minimum divergence between island populations. The populations of P. sonani on the two islands were significantly different in their host ranges, and the genetic clustering and phylogenies of P. sonani established two valid evolutionary species. Integrated species delimitation combining morphological, molecular and ecological characters supported two distinct species of P. sonani from Green Island and Orchid Island. The Green Island population was described as P. jitanasaius sp.n. Chen & Lin, and it is recommended that its threatened conservation status be recognized. Our findings suggest that the inter‐island speciation of endemic organisms inhabiting both islands may be more common than previously thought, and they highlight the possibility that the cryptic diversity of small oceanic islands may still be largely underestimated. 相似文献
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生态风险预警等级评估和演化趋势模拟,可为生态风险管理提供可靠的辅助决策。以重庆市为研究对象,基于驱动力-压力-状态-影响-响应模型,构建重庆市生态风险预警指标体系,采用正态云模型和集对分析法,定量分析重庆市生态风险时空分异特征和演化趋势。研究结果表明:(1)2013-2019年,重庆市生态风险值呈"上升-下降"的波动趋势,综合生态风险隶属于重警等级,生态风险综合值从0.295下降到0.278,生态环境逐年好转;(2)重庆市生态风险有下降、不变、先上升后降低、先降低后上升以及一直升高5种演化趋势,分别占比39%、16%、5%、21%、24%;(3)重庆市生态风险转移分为两个方向,2013-2016年生态风险空间分异性增大,中警、轻警和无警风险等级不断向东北、东南和西部四周分散转移;2016-2019年生态风险分布格局变化较小,重警风险区在东部聚集;(4)演化趋势模拟结果表明,未来重庆市生态风险降低的区县有13个,占比34%,生态环境有向好发展的趋势;生态风险上升的区县有25个,占比66%,生态环境会有所恶化,但是恶化程度较低。将生态风险等级划分与预警演化趋势相结合,能为城市生态风险管理提供科学依据。 相似文献
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