DNA合成提供了驱动力,以加速DNA解的螺旋酶
Natalie M Stano1, Yong-Joo Jeong, Ilker Donmez
1Department of Biochemistry, UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, USA.
Nature
|May 20, 2005
概括
菌体T7螺旋酶缓慢地解开双重DNA. 与T7DNA聚合酶相结合,螺旋酶解速度显著增加,与单链DNA的速度相匹配.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 病毒学 病毒学
背景情况:
- 螺旋酶是分子电机,用于核酸链转位和解的核酸5'-三酸 (NTP) 的水解.
- 菌体T7螺旋酶在单链DNA (ssDNA) 上以130个/秒的速度移动,但在双重DNA上速度减慢了十倍.
研究的目的:
- 调查T7 DNA聚合酶在增强T7螺旋酶双重DNA解速度中的作用.
- 了解DNA聚合酶活动影响基酶转位速度的机制.
主要方法:
- 在体外对T7酶和T7DNA聚合酶活动的表征.
- 测试测量在存在或缺少DNA聚合酶的情况下的DNA解速度.
- 对螺旋酶刺激对DNA合成速率的依赖性的分析.
主要成果:
- T7 DNA聚合酶显著增加了T7螺旋酶解率,达到114个基对/秒.
- 酶刺激取决于DNA合成速率,而不是特定的蛋白质-蛋白质相互作用.
- 高效的双重DNA合成需要化酶和聚合酶的协调作用.
结论:
- T7酶和T7DNA聚合酶的联合作用使得快速的双重DNA解和合成成为可能.
- 通过快速捕获ssDNA的DNA聚合酶活性,通过双重DNA积极驱动螺旋酶转位.
相关概念视频
The DNA Replication Fork
An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
DNA Helicases
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...
The Replisome
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...
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...
DNA Topoisomerases
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. Type I...
Types and Mechanism of action
Topoisomerases are divided into two main types. Type I...
The DNA Replication Fork
An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
The Replisome
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...
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...


