Organization and expression of the human gene for O6-methylguanine-DNA methyltransferase

Yoshimichi Nakatsu, Kazue Hattori, Hiroshi Hayakawa, Kenji Shimizu, Mutsuo Sekiguchi

Research output: Contribution to journalArticle

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Abstract

O6-Methylguanine-DNA methyltransferase plays an important role in cellular defence against mutagens and carcinogens with alkylating activity. Certain tumor-derived cell lines, termed Mer-, are defective in the enzyme activity and have an increased sensitivity to alkylating agents. We cloned the genomic sequence coding for the human O6-methylguanine-DNA methyltransferase and elucidated the structure. The gene consisted of 5 exons and spanned more than 170 kb, while mRNA for the enzyme was 950 nucleotides long. No or only little mRNA for the enzyme was formed in Mer- cells, though there was no gross difference in the coding and promoter regions of the gene between Mer+ and Mer- cells. The putative promoter region, derived from Mer+ cells, was placed upstream of the chloramphenicol acetyltransferase reporter gene and the constructs were introduced into Mer+ and Mer- cells. In Mer- cells, a lowered level of transient expression of the gene was observed as compared with Mer+ cells, but this difference alone does not account for the in vivo difference of expression of the gene in the two types of cells; there might be difference in cis-acting elements. The DNA sequence in the 5′ upstream region of the gene was extremely GC-rich and there were no consensus sequences, such as the TATA and CAAT boxes. There were lower levels of methylation in the putative promoter of various Mer- cells, as compared with findings in Mer+ cells. Methylation in this region may be involved in regulating expression of the gene.

Original languageEnglish
Pages (from-to)119-132
Number of pages14
JournalMutation Research-DNA Repair
Volume293
Issue number2
DOIs
Publication statusPublished - Jan 1993

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Methyltransferases
Genes
Gene Expression
DNA
Methylation
Genetic Promoter Regions
Messenger RNA
Enzymes
Chloramphenicol O-Acetyltransferase
Alkylating Agents
DNA sequences
Mutagens
Enzyme activity
O-(6)-methylguanine
TATA Box
Carcinogens
Tumors
Consensus Sequence
Exons
Tumor Cell Line

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Toxicology
  • Genetics

Cite this

Organization and expression of the human gene for O6-methylguanine-DNA methyltransferase. / Nakatsu, Yoshimichi; Hattori, Kazue; Hayakawa, Hiroshi; Shimizu, Kenji; Sekiguchi, Mutsuo.

In: Mutation Research-DNA Repair, Vol. 293, No. 2, 01.1993, p. 119-132.

Research output: Contribution to journalArticle

Nakatsu, Yoshimichi ; Hattori, Kazue ; Hayakawa, Hiroshi ; Shimizu, Kenji ; Sekiguchi, Mutsuo. / Organization and expression of the human gene for O6-methylguanine-DNA methyltransferase. In: Mutation Research-DNA Repair. 1993 ; Vol. 293, No. 2. pp. 119-132.
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