Cited by 352Cancer-related Molecular PathwaysEpigenetics and DNA MethylationGenomics and Chromatin Dynamics

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The Human ARF Cell Cycle Regulatory Gene Promoter Is a CpG Island Which Can Be Silenced by DNA Methylation and Down-Regulated by Wild-Type p53

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Robertson, Keith D., Jones, Peter A.·1998·Molecular and Cellular Biology

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Abstract

The INK4a/ARF locus encodes two proteins involved in tumor suppression in a manner virtually unique in mammalian cells. Distinct first exons, driven from separate promoters, splice onto a common exon 2 and 3 but utilize different reading frames to produce two completely distinct proteins, both of which play roles in cell cycle control. INK4a, a critical element of the retinoblastoma gene pathway, binds to and inhibits the activities of CDK4 and CDK6, while ARF, a critical element of the p53 pathway, increases the level of functional p53 via interaction with MDM2. Here we clone and characterize the promoter of the human ARF gene and show that it is a CpG island characteristic of a housekeeping gene which contains numerous Sp1 sites. Both ARF and INK4a are coordinately expressed in cells except when their promoter regions become de novo methylated. In one of these situations, ARF transcription could be reactivated by treatment with the DNA methylation inhibitor 5-aza-2'-deoxycytidine, and the reactivation kinetics of ARF and INK4a were found to differ slightly in a cell line in which both genes were silenced by methylation. The ARF promoter was also found to be highly responsive to E2F1 expression, in keeping with previous results at the RNA level. Lastly, transcription from the ARF promoter was down-regulated by wild-type p53 expression, and the magnitude of the effect correlated with the status of the endogenous p53 gene. This finding points to the existence of an autoregulatory feedback loop between p53, MDM2, and ARF, aimed at keeping p53 levels in check.

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Authors: Robertson, K. D., & Jones, P. A.
Affiliations: University of Southern California (US)
Corresponding author: Jones, Peter A.
ORCID: Robertson, Keith D.
Jones, Peter A.
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 1998-11-01
Volume: 18
Issue: 11
Pages: 6457-6473
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.18.11.6457
PMC: PMC109232
OpenAlex: W2115290061
Cited by: 352
References: 79
Citations by year: 2026: 1 · 2024: 2 · 2023: 4 · 2022: 1 · 2021: 3 · 2019: 3 · 2018: 2 · 2017: 5 · 2016: 4 · 2015: 6 · 2014: 5 · 2013: 14 · 2012: 10
FWCI: 15.25
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Molecular and Cellular Biology (publishedVersion)
PubMed Central (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Cancer Institute (R35 CA 49758-09, R35CA049758, T32CA009320, T32 CA 09320-15)
Topics: Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords: ARF genes, INK4a/ARF locus, CpG islands, DNA methylation, promoter silencing, p53, MDM2, Sp1 binding site, E2F1, transcriptional autoregulation, decitabine, tumor suppression, cell cycle regulation
MeSH terms: Base Sequence, Cell Cycle, Cell Line, Cloning, Molecular, Gene Expression Regulation, Humans, Molecular Sequence Data, Promoter Regions, Genetic, Transfection, Down-Regulation, Up-Regulation, Genes, Tumor Suppressor, Genes, p53, Sequence Deletion, Sequence Analysis, DNA, Genes, Reporter, CpG Islands, DNA Methylation, Cyclin-Dependent Kinase Inhibitor p16

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The Human ARF Cell Cycle Regulatory Gene Promoter Is a CpG Island Which Can Be Silenced by DNA Methylation and Down-Regulated by Wild-Type p53

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