一个DNA聚合酶的适应能力 加载器ATPase复合体
Subu Subramanian1, Weilin Zhang2, Siddharth Nimkar1
1Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, TN, USA.
Molecular biology and evolution
|February 1, 2024
概括
在T4-装载器复合体中的突变可以通过改变静电相互作用来恢复功能. 即使是单一的突变也可以弥补显著的功能损失,揭示潜在的适应潜力.
科学领域:
- 分子生物学分子生物学
- 进化生物学 进化生物学
- 生物化学 生物化学
背景情况:
- 在DNA复制过程中,T4紧固件-装载器复合体至关重要.
- 了解突变如何推动分子机器的功能适应是进化研究的关键.
研究的目的:
- 为了研究突变如何能够恢复有缺陷的T4加载器复合体的功能.
- 确定这种复杂系统中适应的基础分子机制.
主要方法:
- 通过域更换创建了一个有缺陷的T4紧固件加载器变体.
- 利用定向进化和深度突变发生的实验.
- 分析了静电相互作用和结合亲缘关系.
主要成果:
- 多个突变通过消除Asp 86和滑动之间的静电排斥来恢复适应性.
- 这表明,最初的适应性下降是由于加载器和的亲和力下降.
- 在DNA近位残留中的补偿突变也改善了适应性.
- 这些发现突出了增加亲和力的潜在能力.
结论:
- 即使是单点突变也可以弥补复杂分子机器中的功能损失.
- 通过优化低于最佳的相互作用来进行适应,从而揭示了进化灵活性.
更多相关视频
07:38DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
Published on: October 6, 2017
14.1K
11:08Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
Published on: June 19, 2018
9.7K
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
The Replisome
33.5K
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...
33.5K
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
DNA Damage can Stall the Cell Cycle
9.2K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
9.2K
Proofreading
6.3K
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...
6.3K
DNA Topoisomerases
31.3K
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. ...
31.3K
