转化Tn10/IS10的三个化学步骤涉及重复使用单个活性位点
1Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Cell
|January 26, 1996
概括
转化酶IS10酶通过三个不同的化学步骤进行DNA转化. 突变揭示了一个单一的活跃部位在转化酶催化所有三个步骤顺序.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 生物化学 生物化学
背景情况:
- 转换是遗传变异的一个关键机制.
- Tn10/IS10转换涉及DNA裂变和链转移.
- 这些反应发生在由转位子末端和转位酶组成的突触复合体内.
研究的目的:
- 为了研究IS10转化酶的催化机制.
- 为了确定不同的活性位点是否催化转化的化学步骤.
- 阐明个体转化酶单体在反应中的作用.
主要方法:
- 在IS10转移酶的位点定向突变发生.
- 在体外生化测试以监测转化步骤.
- 对野生型和突变型转体酶的反应分析.
主要成果:
- 产生了四种转化酶突变,每个突变都取消了特定的化学步骤 (裂变或链转移).
- 突变并没有破坏突触复合体的形成.
- 野生型和突变型转体酶混合物显示,单个单体分子催化裂变和链转移.
结论:
- 在IS10转化酶中,一个单一的活性位点直接催化了非复制性转化的所有三个化学步骤.
- 同一个转化酶单体同时执行DNA裂变和链转移.
- 这些反应由每个转子子末端的一个活性位点单元顺序执行.
相关概念视频
Overview of Transposition and Recombination
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...
DNA-only Transposons
DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
The donor site from where the transposon is excised is either degraded or...
Transfer RNA Synthesis
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
tRNA Activation
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
ATP Synthase: Mechanism
In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased ATP...
tRNA Activation
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...


