| Visualizing the replicating HSV-1 virus using STED super-resolution microscopy |
| Li ZR1,2; Fang C3; Li X1,2; Chen GJ1; Lu DF1,2; Zhou JM[*]1; Su YY3; Liu HM3; Lang FC1
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刊名 | VIROLOGY JOURNAL
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| 2016
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卷号 | 13期号:X页码:e65 |
关键词 | FISH
HSV-1 replication
ICP8
IF
RNA Pol II
STED
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通讯作者 | zhoujm@mail.kiz.ac.cn
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英文摘要 |
BACKGROUND:
Replication of viral genome is the central event during the lytic infectious cycle of herpes simplex virus 1 (HSV-1). However, the details of HSV-1 replication process are still elusive due to the limitations of current molecular and conventional fluorescent microscopy methods. Stimulated emission depletion (STED) microscopy is one of the recently available super-resolution techniques allowing observation at sub-diffraction resolution.
METHODS:
To gain new insight into HSV-1 replication, we used a combination of stimulated emission depletion microscopy, fluorescence in situ hybridization (FISH) and immunofluorescence (IF) to observe the HSV-1 replication process.
RESULTS:
Using two colored probes labeling the same region of HSV-1 genome, the two probes highly correlated in both pre-replication and replicating genomes. In comparison, when probes from different regions were used, the average distance between the two probes increased after the virus enters replication, suggesting that the HSV-1 genome undergoes dynamic structure changes from a compact to a relaxed formation and occupies larger space as it enters replication. Using FISH and IF, viral single strand binding protein ICP8 was seen closely positioned with HSV-1 genome. In contrast, ICP8 and host RNA polymerase II were less related. This result suggests that ICP8 marked regions of DNA replication are spatially separated from regions of active transcription, represented by the elongating form of RNA polymerase II within the viral replication compartments. Comparing HSV-1 genomes at early stage of replication with that in later stage, we also noted overall increases among different values. These results suggest stimulated emission depletion microscopy is capable of investigating events during HSV-1 replication.
CONCLUSION:
1) Replicating HSV-1 genome could be observed by super-resolution microscopy; 2) Viral genome expands spatially during replication; 3) Viral replication and transcription are partitioned into different sub-structures within the replication compartments. |
资助信息 | National Science Foundation of China provided funding to Jumin Zhou under grant number NSFC81471966. Kunming Institute of Zoology, Chinese Academy of Sciences provided funding to Jumin Zhou under grant number Y102421081. Yunnan Provincial Government provided funding to Jumin Zhou under grant numbers 2011HA005 and 2013FA051. Chinese Academy of Sciences provided funding to Jumin Zhou under grant number KSCXZ-EW-BR-6. |
内容类型 | 期刊论文
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源URL | [http://159.226.149.26:8080/handle/152453/9693] |
专题 | 昆明动物研究所_基因调控与表达遗传 昆明动物研究所_动物模型与人类重大疾病机理重点实验室
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作者单位 | 1.Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, NO. 32 Jiaochang Donglu, Kunming, Yunnan 650223, People’s Republic of China 2.University of Chinese Academy of Sciences, Beijing 100049, People’s Republic of China 3.Leica Microsystems Trading Limited, Shanghai 201206, People’s Republic of China
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推荐引用方式 GB/T 7714 |
Li ZR,Fang C,Li X,et al. Visualizing the replicating HSV-1 virus using STED super-resolution microscopy[J]. VIROLOGY JOURNAL,2016,13(X):e65.
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APA |
Li ZR.,Fang C.,Li X.,Chen GJ.,Lu DF.,...&Lang FC.(2016).Visualizing the replicating HSV-1 virus using STED super-resolution microscopy.VIROLOGY JOURNAL,13(X),e65.
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MLA |
Li ZR,et al."Visualizing the replicating HSV-1 virus using STED super-resolution microscopy".VIROLOGY JOURNAL 13.X(2016):e65.
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