通过DMS-MaPseqq检测到的活人细胞中的端粒酶RNA结构异质性
Nicholas M Forino1, Jia Zheng Woo2, Arthur J Zaug3,4
1Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, CA, USA.
Nature communications
|January 22, 2025
概括
大多数人体端粒酶RNA (hTR) 对于端粒酶逆转录酶 (hTERT) 结合而正确折叠. 然而,一些hTR采用非功能性构造,影响细胞中的端粒酶组合和活性.
科学领域:
- 分子生物学分子生物学
- RNA结构和功能 RNA结构和功能
- 生物化学 生物化学
背景情况:
- 人类端粒酶RNA (hTR) 对于端粒酶生物生成至关重要.
- hTR必须折叠成一个特定的多域架构,包括模板伪节点 (t/PK) 和CR4/5域,以结合端粒酶逆转录酶 (hTERT).
- 适当的hTR折叠对于端粒酶活性至关重要.
研究的目的:
- 为了研究活体细胞内hTR的体内折叠.
- 确定采用功能性与非功能性形状的hTR的比例.
- 评估替代hTR构造对端粒酶组合和活性的影响.
主要方法:
- 使用二甲基硫酸盐突变分析与测序 (DMS-MaPseq) 来探测活细胞中的hTR结构.
- 综合解卷分析被用来解释结构数据.
- 对净化端粒酶RNP复合物的分析进行了比较.
主要成果:
- 大约15%的细胞hTR存在于CR4/5形状,防止hTERT结合.
- 这种错误折叠的hTR的比例是独立于hTERT表达水平的.
- 稳定替代CR4/5形状的突变会损害端粒酶的组合和活性.
- 在纯化端粒酶RNP复合体中没有检测到替代CR4/5形状.
结论:
- 细胞hTR的很大一部分可能存在于非功能性构造中,可能充当动力折叠陷.
- 只有hTR的初级CR4/5适合器似乎在端粒酶组合中活跃.
- 这种细胞错误折叠表明了端粒酶活性的调节机制,并突出了体内RNA折叠的挑战.
更多相关视频
08:26Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein
Published on: June 12, 2018
9.8K
05:412D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
Published on: July 10, 2020
1.9K
相关概念视频
02:41Telomeres and Telomerase
23.0K
In eukaryotic DNA replication, a single-stranded DNA fragment remains at the end of a chromosome after the removal of the final primer. This section of DNA cannot be replicated in the same manner as the rest of the strand because there is no 3’ end to which the newly synthesized DNA can attach. This non-replicated fragment results in gradual loss of the chromosomal DNA during each cell duplication. Additionally, it can induce a DNA damage response by enzymes that recognize single-stranded...
23.0K
03:21RNA-seq
9.8K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.8K
02:31Replication in Eukaryotes
170.2K
Overview
170.2K