在增殖皮细胞中追踪复制HPV基因组
Jonathan R Shin1, Ivan Avilov2, Mario Schelhaas2
1Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
bioRxiv : the preprint server for biology
|April 1, 2025
概括
研究人员使用ANCHOR技术开发了一种新方法来追踪活细胞中人类乳头瘤病毒 (HPV) DNA. 这允许实时监测HPV18基因组复制和在角质细胞中的分布.
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:
- 人类乳头瘤病毒 (HPV) 基因组在持续感染期间充当小染色体.
- 在活细胞中监测病毒基因组动态是具有挑战性的,因为基因组的小尺寸和紧的结构.
研究的目的:
- 开发一种使用ANCHOR技术在活细胞中检测和跟踪HPV18基因组的新方法.
- 为了可视化HPV18基因组的复制,维护和分割在增殖的角质细胞中.
主要方法:
- 将ANCHOR cis元素纳入HPV18基因组.
- 从含有病毒源和增强剂的HPV18依赖复制体中表达ParB-GFP蛋白.
- 利用一种neomycin耐药基因在细菌和真核细胞中进行选择.
- 实时跟踪在活的角质细胞中光标记的HPV18基因组.
主要成果:
- 通过光蛋白检测,ANCHOR技术使HPV18基因组的间接监测成为可能.
- 复制HPV18-ANCH3基因组在增殖的角质细胞内实时跟踪.
- 观察到HPV18基因组在细胞分裂过程中通过随机连接到宿主染色体,与子细胞相对均地分裂.
结论:
- 开发的分子工具有助于研究活细胞中的HPV基因组动态.
- 这种方法为HPV基因组在感染期间的维护,结合和放大提供了洞察力.
- 这些发现阐明了HPV18DNA在分裂宿主细胞中的分区机制.
更多相关视频
13:41Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
12.3K
10:26RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma
Published on: March 11, 2014
27.2K
相关概念视频
02:15Replicative Cell Senescence
3.6K
Replicative cell senescence is a property of cells that allows them to divide a finite number of times throughout the organism's lifespan while preventing excessive proliferation. Replicative senescence is associated with the gradual loss of the telomere — short, repetitive DNA sequences found at the end of the chromosomes. Telomeres are bound by a group of proteins to form a protective cap on the ends of chromosomes. Embryonic stem cells express telomerase — an enzyme that adds...
3.6K
01:29Replication in Eukaryotes
12.8K
In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...
12.8K
02:37Restarting Stalled Replication Forks
5.7K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
5.7K
02:31Chromosome Replication
8.6K
Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins. This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin...
8.6K