相关实验视频
Updated: Jul 23, 2025

06:48
CD Spectroscopy to Study DNA-Protein Interactions
Published on: February 10, 2022
6.8K
通过双链DNA脱氨酶毒素 DddAddA的序列特异性细胞酸脱氨的结构基础
Lulu Yin1,2,3, Ke Shi1,2,3, Hideki Aihara4,5,6
1Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN, USA.
Nature structural & molecular biology
|July 17, 2023
概括
细菌间除氨酶毒素DddA通过挤出细胞因子来编辑DNA,避免基因翻转. 这种由晶体结构揭示的机制为基因组编辑工具提供了新的可能性.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 细菌间的除氨酶毒素DddA在双链DNA中进行细胞酸转化为尿素.
- DddA促进了无CRISPR的线粒体基编辑,但其基质选择性机制尚不清楚.
研究的目的:
- 阐明DddA基质选择性的分子机制.
- 确定DddA的DNA编辑活动的结构基础.
主要方法:
- 进行X射线晶体学以获得DddA-dsDNA复杂结构.
- 生物化学测试以评估除氨酶活性和序列选择性.
主要成果:
- 晶体结构显示,DddA在小槽中结合了dsDNA,挤出了目标细胞蛋白.
- 确定了一种涉及Phe1375的双重排位机制,其中胺取代了细胞因子.
- 基因基因不匹配增强了DddA活动,并降低了序列的特异性.
- 确定了具有改变活动或基质偏好的突变物.
结论:
- DddA 串联移位机制允许在没有基翻转的情况下进行细胞蛋白编辑.
- 了解DddA的机制可以为新型基因组编辑工具的设计提供信息.
- DddA的活性是由DNA序列背景和不匹配调节的.
相关概念视频
Maxam-Gilbert Sequencing
11.3K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
11.3K
Overview of DNA Repair
31.1K
In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
Chemically...
Chemically...
31.1K
DNA Base Pairing
27.6K
Erwin Chargaff’s rules on DNA equivalence paved the way for the discovery of base pairing in DNA. Chargaff’s rules state that in a double-stranded DNA molecule,
27.6K
Fixing Double-strand Breaks
12.7K
The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
12.7K
Mismatch Repair
4.9K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.9K
Base Excision Repair
22.6K
One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
The first step of...
22.6K

