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Updated: Oct 9, 2026

Yeast As a Chassis for Developing Functional Assays to Study Human P53
14:57

Yeast As a Chassis for Developing Functional Assays to Study Human P53

Published on: August 4, 2019

A promoter function of the CCCGGG Sma I recognition sequence and its specific role in determining p53 status and

A L Yang1, M F Festing

  • 1MRC Toxicology Unit, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom. aly1@le.ac.uk

Biochemical and Biophysical Research Communications
|February 22, 2001
PubMed

Insights

Researchers identified the CCCGGG sequence as a transcriptionally active DNA-binding site for the tumor suppressor p53 protein. This finding enables the pGL3-Basic vector to indicate p53 activity and identify p53-regulated genes.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cancer Research

Background:

  • The tumor suppressor p53 protein is crucial for cellular responses to DNA damage.
  • A common in vivo DNA-binding site for p53 has remained unidentified.
  • The pGL3-Basic vector is designed for studying regulatory sequences without endogenous promoters.

Purpose of the Study:

  • To identify a transcriptionally active DNA-binding site for p53.
  • To evaluate the pGL3-Basic vector's potential for assessing p53 activity.
  • To explore the CCCGGG sequence as a potential p53 consensus site.

Main Methods:

  • Electrophoretic mobility shift assay (EMSA) to assess p53 binding to DNA in vitro.
  • Utilizing the pGL3-Basic vector with its CCCGGG sequence to monitor p53-dependent transcription.
  • Analyzing the presence of the CCCGGG sequence in known p53-regulated genes.

Main Results:

  • The CCCGGG sequence (Sma I site) in the pGL3-Basic vector promoted p53-dependent transcription of the luciferase gene.
  • Human p53 demonstrated in vitro binding to the CCCGGG sequence via EMSA.
  • The CCCGGG sequence was identified in promoter regions of several p53-regulated genes.

Conclusions:

  • The CCCGGG sequence is the first identified transcriptionally active DNA-binding site for p53.
  • The pGL3-Basic vector can serve as an indicator of p53 transcriptional activity.
  • The CCCGGG sequence is proposed as a consensus site for p53 recognition in vivo, aiding in the identification of p53-controlled genes.

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