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DNA Helicases00:55

DNA Helicases

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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...
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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 DNA Replication Fork01:02

The DNA Replication Fork

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DNA Replication02:40

DNA Replication

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DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication...
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The Replisome03:01

The Replisome

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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.
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...
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The Replisome03:01

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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水素結合なしのDNA複製に関する構造的洞察

Karin Betz1, Denis A Malyshev, Thomas Lavergne

  • 1Departments of Chemistry and Biology, Konstanz Research School Chemical Biology, Universität Konstanz , Universitätsstrasse 10, D-78464 Konstanz, Germany.

Journal of the American Chemical Society
|November 29, 2013
PubMed
まとめ
この要約は機械生成です。

d5SICS-dNaMのような不自然な塩基対で遺伝子アルファベットを拡張すると,DNAの潜在能力を高めます. しかし,それらの複製は,ポリメラーゼの相互作用と塩基対のインターキャレーションによる課題に直面しています.

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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
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科学分野:

  • 分子生物学は分子生物学である.
  • 合成生物学 合成生物学とは
  • バイオケミストリー バイオケミストリー

背景:

  • 不自然な塩基対 (UBP) による遺伝アルファベットの拡張は,遺伝的および化学的潜在力の増加を約束する.
  • d5SICS-dNaMペアは,効率的に複製されるUBPですが,その複製メカニズムは,自由DNA内のインターカレート構造のために不明のままです.

研究 の 目的:

  • d5SICS.の反対側にdNaMTPを挿入する際に生じる化学前および化学後複合体の形成を調査する.
  • d5SICS-dNaM不自然な塩基対の複製を制御する構造的ダイナミクスとポリメラーゼ相互作用を解明する.

主な方法:

  • d5SICS.の反対のdNaMTP挿入のためのプレケミストリー複合体の特徴.
  • 不自然な塩基対の位置と挿入後の形状を詳細に記述するポスト化学複合体の分析.
  • ポリメラーゼ活性部位の相互作用と核酸結合親和性を調査する.

主要な成果:

  • d5SICSTP挿入とは異なり,dNaMTP添加は完全に閉じたポリメラーゼ状態を誘導しません.
  • 挿入後,d5SICS-dNaMペアは,2つの異なるモードを通じてポリメラーゼ活性部位内で相互に作用し,側面の核酸によって影響を受けます.
  • インターケレーションは,次の正しいトリフォスファート結合に対する親和性を低下させ,プライマーのさらなる拡張を妨げます.

結論:

  • d5SICS-dNaMの非自然な塩基対の複製は,挿入後のインターケレーションと,その後,ディインターケレーションとアクティブサイト再配置の必要性によって制限されています.
  • これらの構造的ダイナミクスを理解することは,UBPの複製を最適化し,合成生物学を前進させるために不可欠です.