相关实验视频
Updated: Aug 17, 2026

08:40
Methyl-binding DNA capture Sequencing for Patient Tissues
Published on: October 31, 2016
通过c-Myc基本区域的甲基化敏感序列特定DNA结合
1Howard Hughes Medical Institute, Department of Biochemistry, New York, NY.
概括
研究人员探索了c-Myccoprotein在细胞生长中的功能. 他们发现c-Myc基本区域与DNA结合,影响细胞生长,并在细胞转化中与H-ras合作.
科学领域:
- 分子生物学分子生物学
- 在瘤学瘤学.
- 细胞生物学 细胞生物学
背景情况:
- 在调节细胞生长方面,c-Myccoprotein的精确功能在很大程度上仍未确定.
- 在结构上,c-Myc 具有与螺旋环螺旋 (HLH) 和氨酸拉链蛋白质中发现的 DNA 结合基因相似的基本区域.
研究的目的:
- 为了研究c-Myc基本区域的DNA结合能力.
- 确定c-Myc的DNA结合特异性是否与其在细胞生长控制和瘤发生中的作用有关.
主要方法:
- 通过将c-Myc基本区域与E12增强剂结合因子的HLH二元化域融合,设计了一种嵌合蛋白 (E6).
- 使用电泳运动转移试验 (EMSA) 来评估E6对E盒元素的DNA结合活性.
- 使用甲基化干扰试验来探测E6结合的序列特异性.
- 进行了一项大鼠胚胎纤维细胞转化试验,以评估与H-ras.合作的E6表达的功能后果.
主要成果:
- 嵌合体E6蛋白以一种取决于c-Myc基本区域完整性的方式特别与E盒DNA元素结合.
- 在E盒内CpG位点的甲基化特别抑制了E6结合,表明了序列特定的识别.
- 功能性E6的表达,但不是DNA结合突变,抑制了由c-myc和H-ras.由老鼠胚胎纤维细胞的合作转化.
结论:
- 该c-Myc基本区域具有固有的DNA结合活性,具有特定的识别特性.
- c-Myc的DNA结合功能与其在细胞转化中与其他癌基因 (如H-ras) 合作的能力有关.
- 这些发现提供了关于c-Myc在细胞生长和癌症中的作用背后的分子机制的见解.
相关概念视频
Epigenetic Regulation
Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
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,...
Cis-regulatory Sequences
Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
Epigenetic Regulation
Epigenetic changes alter the physical structure of the DNA without changing the genetic sequence and often regulate whether genes are turned on or off. This regulation ensures that each cell produces only proteins necessary for its function. For example, proteins that promote bone growth are not produced in muscle cells. Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
X-chromosome...
X-chromosome...
Induced Pluripotent Stem Cells
Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic cells are...
Somatic cells are...

