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相关概念视频

The DNA Replication Fork01:02

The DNA Replication Fork

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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...
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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of...
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Restarting Stalled Replication Forks02:37

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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,...
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Replication in Prokaryotes01:32

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DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
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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.
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The Replisome03:01

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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.
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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在蛋白相互作用选中发现了DONSON在复制启动中的作用

Yang Lim1, Lukas Tamayo-Orrego2, Ernst Schmid1

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA.

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概括

通过组装CDC45-MCM2-7-GINS (CMG) 螺旋酶,DONSON蛋白对于真核DNA复制的启动至关重要. 突变导致小脑化, 凸显了它在人类发展中的作用.

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科学领域:

  • 分子生物学
  • 遗传学
  • 生物化学

背景情况:

  • 细胞DNA复制的启动依赖于CDC45-MCM2-7-GINS (CMG) 螺旋酶的组合.
  • 在酵母中,一个预加载复合体 (pre-LC) 促进了GINS与染色质相关的MCM2-7结合,形成CMG.
  • 脊椎动物中CMG组装的确切机制尚不完全理解.

研究的目的:

  • 研究甲状动物蛋白DONSON在脊椎动物中CMG酶组合中的作用.
  • 在复制启动的背景下阐明DONSON的功能及其与小脑原始矮体的潜在联系.

主要方法:

  • 使用AlphaFold进行蛋白质与蛋白质相互作用的选.
  • 进行预测蛋白相互作用的实验验证.
  • 在小鼠模型中研究了患者衍生的DONSON突变对CMG组合及其表型后果的影响.

主要成果:

  • DONSON作为脊椎动物前LC的支架,包括GINS,TOPBP1和DNA聚合酶epsilon.
  • DONSON促进了前LC的对接到MCM2-7,从而为CMG形成提供了GINS.
  • 一个来自患者的DONSON突变损害了CMG组合,并在小鼠中引起小脑化.

结论:

  • DONSON对于脊椎动物的CMG酶组合至关重要,统一了对真核生物复制启动的理解.
  • 由于DONSON突变导致的CMG组合缺陷与小脑有关.
  • 在蛋白蛋白相互作用查中,以AlphaFold为例,加速了机械生物学发现.