崩状态调解了DNA聚合酶β I260突变的低忠度
Carel Fijen1, Cristian Chavira2,3, Khadijeh Alnajjar1,2
1Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-8034, United States.
Biochemistry
|September 19, 2024
概括
在DNA聚合酶β (Pol β) 中的I260 M突变降低了DNA修复中的精度,可能导致癌症. 这种突变导致"崩"的蛋白质状态,损害了DNA合成期间的核酸选择.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- DNA聚合酶β (Pol β) 对于基切除修复至关重要,填补单核酸缺口.
- 聚β缺乏与突变发生的增加,基因组不稳定性和癌症有关.
- 在前列腺癌中发现的Pol β中的I260 M体质突变表现出减少的核酸歧视.
研究的目的:
- 阐明I260 M Pol β突变体降低保真度背后的分子机制.
- 调查I260 M突变如何影响核酸结合和构造变化.
主要方法:
- 单个周转动力学研究核酸结合.
- 稳态光和福斯特共振能量转移 (FRET) 用于分析形状变化.
- 动力建模用于比较野生型 (WT) 和I260 M Pol β.
主要成果:
- I260 M 突变会在 Pol β. 的手指区域产生"崩"状态.
- I260 M表现出对温度敏感的突变活性和比WT更紧密的核酸结合.
- 与WT相比,I260 M的形状变化中缺少一个关键的前催化非共价步骤.
结论:
- I260 M Pol β 的"崩"状态损害了核酸的区分,降低了聚合酶的忠实性.
- "指关闭"的形状变化对于保持聚合酶忠实性和精确的DNA合成至关重要.
- 了解这些机制突出了Pol β忠实性在预防癌症中的作用.
相关概念视频
Translesion DNA Polymerases
9.9K
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...
9.9K
Proofreading
6.2K
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...
6.2K
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 Topoisomerases
31.1K
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. ...
31.1K
Homologous Recombination
50.3K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.3K
The Replisome
33.2K
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...
33.2K


