Related Experiment Video

Updated: Aug 21, 2026

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
06:07

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Published on: August 5, 2022

Regulation of human methylthioadenosine phosphorylase gene by the CBF (CCAAT binding factor)/NF-Y (nuclear factor-Y)

Yuwaraj Kadariya1, Kaname Nakatani, Junji Nishioka

  • 1Department of Laboratory Medicine, Mie University School of Medicine, Tsu, Mie 514-8507, Japan. yuwaraj.kadariya@fccc.edu

The Biochemical Journal
|November 13, 2004
PubMed

Insights

Human 5'-deoxy-5'-methylthioadenosine phosphorylase (hMTAP) gene expression is regulated by CCAAT-binding factor (CBF). The distal CCAAT motif significantly enhances hMTAP transcriptional activation, crucial for understanding its role in cancer.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Human 5'-deoxy-5'-methylthioadenosine phosphorylase (hMTAP) is vital in the methionine salvage pathway.
  • hMTAP inactivation is common in human tumor cells, highlighting the need to understand its gene regulation.

Purpose of the Study:

  • To elucidate the transcriptional regulation mechanism of the hMTAP gene.
  • To identify cis-acting regulatory elements and transcription factors involved in hMTAP gene expression.

Main Methods:

  • Cloning and functional analysis of the hMTAP promoter using luciferase reporter assays.
  • Electrophoretic mobility-shift assays (EMSA) and supershift assays to identify DNA-protein interactions.
  • Chromatin immunoprecipitation (ChIP) assays to confirm in vivo binding of transcription factors.

Main Results:

  • Maximal hMTAP promoter activity was observed between -446 and -152, containing two CCAAT elements.
  • CCAAT-binding factor (CBF), also known as nuclear factor-Y, binds to both proximal and distal CCAAT motifs.
  • The distal CCAAT motif exhibits stronger CBF binding and plays a major role in transcriptional activation.
  • Seven distinct transcriptional start sites were identified between -135 and -58.

Conclusions:

  • hMTAP gene expression is transcriptionally regulated by CBF.
  • The distal CCAAT motif is a key regulatory element for hMTAP transcriptional activation.
  • Understanding hMTAP regulation provides insights into its role in cancer biology.

Related Concept Videos

Master Transcription Regulators02:23

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 Regulation01:46

Epigenetic Regulation

Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
Epigenetic Regulation01:37

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...
Biosynthesis of Nucleic Acids01:28

Biosynthesis of Nucleic Acids

Nucleic acid biosynthesis is a fundamental biochemical process that produces the purine and pyrimidine nucleotides essential for DNA and RNA synthesis. This pathway maintains a balanced nucleotide pool, preventing imbalances that could jeopardize genetic integrity and cellular function. Given the crucial role of nucleotides, their synthesis is tightly regulated to ensure proper cellular homeostasis.Purine BiosynthesisThe biosynthesis of purine nucleotides begins with ribose-5-phosphate, a...
General Transcription Factors01:30

General Transcription Factors

Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...