サイト固有の再結合シナプスのDNAと複合したCre再結合酵素の構造
関連する実験動画
Updated: Nov 1, 2026
07:06
Principles of Site-Specific Recombinase (SSR) Technology
Published on: May 29, 2008
F Guo1, D N Gopaul, G D van Duyne
1Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Nature
|September 4, 1997
まとめ
バクテリオファグのCreリコンビネーゼは,ロックスP部位を結合することにより,遺伝的再編成を促進します. その結晶構造は,反応中間物質を明らかにし,現存するサイト固有の再結合のモデルに挑戦しています.
科学分野:
- 分子生物学は分子生物学である.
- 構造生物学 構造生物学とは
- 遺伝学 遺伝学とは
背景:
- サイト固有のDNA再結合は,ウイルス統合,切除,染色体分離に不可欠な遺伝的再編成を可能にします.
- リコンビナーゼ酵素は,特定のDNA配列を認識し,鎖交換を触媒化することで,このプロセスを媒介する.
研究 の 目的:
- バクテリオファージの構造的メカニズムを解明するために,Creリコンビネーゼ媒介サイト特異再結合.
- クレ-ロックスPの再結合で形成される中間物質の構造的基礎を調査する.
主な方法:
- 2.4 Å解像度でLoxPDNA基板に結合するCreリコンビネーゼの構造を決定するために,X線結晶学を用いた.
- 構造分析は,再結合反応中の主要な中間物質と構成の変化を特定することに焦点を当てました.
主要な成果:
- 結晶構造は,クレア分子が分裂したDNA基板と共振性3'-ホスフォチロジン結合を形成する反応中間物質を明らかにした.
- 4つの再結合と2つのロックスP部位を含むシナプス構造が観察され,ホリデイ-ジャンクション中間物質に似ている.
- 観測された構造は,重要な四次構造の変化を必要とするモデルに挑戦し,微妙なアロステリックシフトが反応を調整することを示唆しています.
結論:
- Cre-loxP複合体の構造は,分子レベルでのサイト固有の再結合のメカニズムについての洞察を提供します.
- クレレ・リコンビネーゼにおける微妙なアロステル変化は,DNAの分裂と鎖交換を調節し,代替的なメカニズムモデルを提供することができる.
関連する概念動画
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
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02:31Homologous Recombination
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...
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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...
03:01The 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...
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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...