Identification of the functional elements in the promoter region of human DNA topoisomerase IIIβ gene

Young Hoon Cho, Jee Young Park, Sang Youp Han, In Kwon Chung

Research output: Contribution to journalArticle

Abstract

In this study, we have isolated and characterized the promoter region of the human DNA topoisomerase IIIβ (hTOP3β) gene. The 5′ RACE assay showed a short exon 1 encoding only the 35-bp untranslated region and suggested the presence of multiple transcription initiation sites. The hTOP3β gene promoter lacks a canonical TATA box or initiation element and is moderately high in GC content. Transient expression of a luciferase reporter gene under the control of serially deleted 5′-flanking sequence identified an activator element between -141 and -119 upstream of the transcription initiation site and a second regulatory element between -91 and -71. On the basis of scanning mutations of triple nucleotides, we demonstrated that a 5′GGAACC3′ element between -117 and -112 plays a critical role in the up-regulation of the basal transcription activity. Changing the 5′GGAACC3′ sequence leads to markedly reduced promoter activity. Gel mobility shift assays revealed that the 5′GGAACC3′ element is required for DNA binding by the transcription factor complex. These observations lead to the conclusion that the positive regulatory region including the 5′GGAACC3′ core element is essential for efficient expression of the hTOP3β gene as well as for the binding of as yet unidentified regulatory factor(s).

Original languageEnglish
Pages (from-to)272-278
Number of pages7
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1679
Issue number3
DOIs
Publication statusPublished - 2004 Sep 17

Fingerprint

Transcription Initiation Site
Genetic Promoter Regions
Genes
Untranslated Regions
TATA Box
5' Flanking Region
Nucleic Acid Regulatory Sequences
Base Composition
Electrophoretic Mobility Shift Assay
Luciferases
Reporter Genes
Exons
Assays
Transcription Factors
Up-Regulation
Nucleotides
Gels
Gene Expression
Mutation
DNA

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Genetics

Cite this

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title = "Identification of the functional elements in the promoter region of human DNA topoisomerase IIIβ gene",
abstract = "In this study, we have isolated and characterized the promoter region of the human DNA topoisomerase IIIβ (hTOP3β) gene. The 5′ RACE assay showed a short exon 1 encoding only the 35-bp untranslated region and suggested the presence of multiple transcription initiation sites. The hTOP3β gene promoter lacks a canonical TATA box or initiation element and is moderately high in GC content. Transient expression of a luciferase reporter gene under the control of serially deleted 5′-flanking sequence identified an activator element between -141 and -119 upstream of the transcription initiation site and a second regulatory element between -91 and -71. On the basis of scanning mutations of triple nucleotides, we demonstrated that a 5′GGAACC3′ element between -117 and -112 plays a critical role in the up-regulation of the basal transcription activity. Changing the 5′GGAACC3′ sequence leads to markedly reduced promoter activity. Gel mobility shift assays revealed that the 5′GGAACC3′ element is required for DNA binding by the transcription factor complex. These observations lead to the conclusion that the positive regulatory region including the 5′GGAACC3′ core element is essential for efficient expression of the hTOP3β gene as well as for the binding of as yet unidentified regulatory factor(s).",
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Identification of the functional elements in the promoter region of human DNA topoisomerase IIIβ gene. / Cho, Young Hoon; Park, Jee Young; Han, Sang Youp; Chung, In Kwon.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1679, No. 3, 17.09.2004, p. 272-278.

Research output: Contribution to journalArticle

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