在晶体中观察DNA聚合酶中的活性位点动态和过渡性金属离子结合
Caleb Chang1, Grace Zhou1, Yang Gao1
1Department of Biosciences, Rice University, Houston, Texas 77005, USA.
Structural dynamics (Melville, N.Y.)
|June 19, 2023
概括
DNA聚合酶使用双金属离子机制进行DNA合成和修复. 时间分辨率晶体学揭示了瞬态,原料对齐和金属离子在催化和分辨中的作用.
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:
- DNA聚合酶是DNA复制和修复的必需酶.
- 之前的研究通过动力学研究和静态X射线晶体学阐明了一般的运动路径和一个双金属离子催化机制.
研究的目的:
- 审查和整合来自DNA聚合酶的静态和时间解析结构数据的见解.
- 突出了原料对齐和金属离子相互作用在DNA聚合酶功能中的重要性.
主要方法:
- 关于DNA聚合酶结构的现有文献的审查.
- 分析基于扩散的时间解析晶体学数据.
- 静态和动态结构信息的比较.
主要成果:
- 时间分辨率晶体学可视化了以原子分辨率的催化反应.
- 过渡事件和金属离子结合,以前难以捉摸,现在可以观察到.
- 结构数据强调了原料对齐在催化中的关键作用.
- 不同的金属离子结合事件对于基质歧视至关重要.
结论:
- 动态结构研究为DNA聚合酶机制提供了前所未有的见解.
- 原料的对齐和特定的金属离子协调是聚合酶忠实性和活性的关键决定因素.
- 了解这些机制对于开发针对DNA复制和修复的治疗策略至关重要.
相关概念视频
Translesion DNA Polymerases
10.0K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
10.0K
Proofreading
6.4K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
Errors During Replication are Corrected by the DNA Polymerase...
6.4K
The Replisome
34.0K
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
34.0K
DNA Topoisomerases
31.5K
Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
31.5K
DNA Helicases
21.6K
DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
21.6K
Restarting Stalled Replication Forks
5.8K
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,...
5.8K


