ガンマ・デルタ・リゾルバースの変異体:再結合機能の遺伝子解析

B J Newman, N D Grindley

Cell

|September 1, 1984

まとめ

DNA再結合を理解するために,レゾルバゼタンパク質変異体が研究されました. コインテグレート解像度に影響する特定の変異は,解像度アミノ端末領域の保存された領域で特定されました.

科学分野:

分子生物学は分子生物学である.
遺伝学遺伝学とは
バイオケミストリーバイオケミストリー

背景:

ガンマデルタトランポゾンは,サイト固有の再結合と遺伝子調節という二重の機能を持つ溶解酵素タンパク質をコードする.
両方のリゾルバース機能は,リゾルバースサイトとの相互作用に依存しています.

研究の目的:

DNA再結合反応にのみ関与する溶解酵素タンパク質の特定の領域を特定する.
再結合活性を維持しながら,共統合解像度に欠陥のある突然変異体を特徴付けるため.

主な方法:

独立レズルバース変異体の分離と特徴付け.
規制活動を通じてリゾートサイトへの突然変異の結合を評価する.
ミュータントタンパク質におけるアミノ酸置換分析.

主要な成果:

6つの異なるアミノ酸置換を持つ9つの変異体が特定されました.
すべての置換は,resolvaseのN端140アミノ酸内で発生しました.
変異は2つの保存領域に群集し,機能領域を示唆した.

結論:

N末端領域内の特定のアミノ酸残基は,再結合におけるリゾルバゼの触媒活性にとって重要である.
1つの保存されたクラスタは,再結合中に触媒ドメインとクロスオーバーサイト間の相互作用を媒介する可能性があります.

さらに関連する動画

11:40

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

Published on: June 25, 2013

09:27

Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins

Published on: January 17, 2014

関連する概念動画

00:57

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.

02:31

Homologous 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...

02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

02:53

Conservative 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...

02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon has three reading...