相关实验视频
Updated: Jul 29, 2026

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
对依赖RNA的RNA聚合酶网格的可视化和功能分析
John M Lyle1, Esther Bullitt, Kurt Bienz
1Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
概括
病毒RNA复制利用宿主细胞膜进行. 脊髓灰质炎病毒RNA聚合酶在这些膜上形成2D阵列,增强RNA结合和复制效率.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 阳性链RNA病毒,如脊髓灰质炎病毒,在宿主细胞内复制它们的遗传物质,特别是在细胞内膜上.
- 脊髓灰质炎病毒RNA依赖性RNA聚合酶对于病毒基因组复制至关重要.
研究的目的:
- 为了研究在复制过程中脊髓灰质炎病毒RNA依赖RNA聚合酶的结构组织.
- 探索宿主细胞内膜在病毒RNA合成中的作用.
主要方法:
- 电子显微镜被用于可视化净化脊髓灰质炎病毒RNA依赖RNA聚合酶的结构.
- 分析影响聚合酶结构和功能的突变.
- 从受感染细胞中分离和检查膜状囊泡.
主要成果:
- 纯化脊髓灰质炎病毒RNA依赖RNA聚合酶形成平面和管状的寡合组.
- 这些聚合酶阵列的完整性对于合作RNA结合和延长至关重要.
- 在聚合酶内部破坏分子间接触的突变会影响阵列结构和功能.
- 在受感染细胞的囊泡表面观察到二维聚合酶阵列.
结论:
- 主体细胞质膜可能作为二维RNA-依赖RNA聚合酶数组的支架.
- 这种与膜相关的组织可以通过表面催化增强病毒RNA复制.
相关概念视频
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
RNA Polymerase II Accessory Proteins
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
Transcription Initiation
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
The promoters and enhancers and their accessory proteins allow tight regulation of...
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...

