在体内与甲对蛋白质-DNA相互作用进行映射:证据表明,在高度转录的基因上保留了组素H4的证据
M J Solomon1, P L Larsen, A Varshavsky
1Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Cell
|June 17, 1988
概括
这项研究表明,即使在活跃的转录过程中,histon H4 仍然在体内与热冲击蛋白70 (hsp70) 基因结合. 甲交叉连接技术在体内绘制了蛋白质-DNA相互作用和染色质结构.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 染色体生物学 染色体生物学
背景情况:
- 了解体内染色体结构对于基因调节至关重要.
- 之前的体外研究表明,在活性转录过程中可以去除组织蛋白,但体内证据缺乏.
研究的目的:
- 为了研究Drosophila melanogaster热冲击蛋白70 (hsp70) 基因的体内染色质结构.
- 为了确定基因组是否仍然与活跃转录的基因有关.
主要方法:
- 在完整的细胞内通过甲介导的蛋白质-DNA交联.
- 在体内交叉连接的染色质碎片的免疫沉.
- 分析hsp70基因染色质结构和基因素结合.
主要成果:
- 热冲击诱导改变了hsp70基因染色质结构,减少了体内蛋白质-DNA接触.
- 基因素H4在体内仍然与hsp70DNA结合,无论转录活动如何.
- 基于甲的映射技术是有效的探测体内蛋白质-DNA相互作用.
结论:
- 在活体中活跃转录的基因中,基因组不一定缺席.
- 甲交叉连接为研究体内染色质动力学和蛋白质-DNA相互作用提供了有价值的工具.
- 该方法可以绘制蛋白质-DNA相互作用的地图,并确定全基因组的染色体蛋白质位置.
相关概念视频
The Nucleosome Core Particle
Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
The paradox
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...
The paradox
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...
Histone Modification
The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression.
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
Spreading of Chromatin Modifications
The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer is an enzyme that can...
Writers
The writer is an enzyme that can...
Euchromatin
The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions take up more dye, appearing darker, while the less-compact areas take up less dye and appear lighter. Based on the compaction level, chromatins are classified into two primary forms – euchromatin and heterochromatin.
Euchromatin is the less dense region of the chromatin and stains lighter. Euchromatin contains histone H3 extensively...
Euchromatin is the less dense region of the chromatin and stains lighter. Euchromatin contains histone H3 extensively...
Heterochromatin
The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions that take up more dye are called heterochromatin. Heterochromatin is further classified into two forms – constitutive heterochromatin and facultative heterochromatin.
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at 9th...
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at 9th...
The Nucleosome Core Particle
Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...


