Molecular mechanisms of human single-minded 2 (SIM2) gene expression: Identification of a promoter site in the SIM2 genomic sequence

Akiko Yamaki, Junko Tochigi, Jun Kudo, Shinsei Minoshima, Nobuyoshi Shimizu, Yoshiko Shimizu

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We previously postulated that the single-minded 2 (SIM2) gene identified on the human chromosome 21q22.2 is a good candidate gene for the pathogenesis of mental retardation in Down syndrome because its mouse homolog exhibits preferential expression in the mouse diencephalon during early embryogenesis. We analyzed the genomic sequence of the entire SIM2 gene which consists of 11 exons and spans over 50 kb. As a step toward understanding the molecular mechanisms of SIM2 gene expression, we have analyzed the human SIM2 gene expression in nine established human cell lines. Three transcripts of 3.6, 4.4, and 6.0 kb were detected in the glioblastoma cell line, T98G, neuroblastoma cell line, TGW, and transformed embryonic kidney cell line, 293. The RACE analysis using SIM2-expressing human cell line T98G provided evidence for the transcription start site at ∼1.2 kb upstream of the translation initiation site. The transfection assay using various deletion constructs with reporter gene suggested the presence of a presumptive promoter region. Transient transfection assay in T98G cell line revealed a significant promoter activity located in the 60 bp sequence between nt -1385 and -1325 upstream region of the translation initiation site. This 60 bp sequence contains cis-elements for c-myb, E47 and E2F transcription factors. Moreover, the gel retardation assay using oligo-DNA of various cis-element sequences indicated the presence of protein factor(s) which bind to the cis-element for c-myb. These results suggested that binding of a protein transcription factor(s) such as c-myb or that alike regulates transcription of the SIM2 gene by binding to a small upstream region.

Original languageEnglish
Pages (from-to)265-275
Number of pages11
JournalGene
Volume270
Issue number1-2
DOIs
Publication statusPublished - 2001 May 30

Fingerprint

Gene Expression
Cell Line
Genes
Transfection
E2F Transcription Factors
Diencephalon
Transformed Cell Line
Transcription Initiation Site
Human Chromosomes
Electrophoretic Mobility Shift Assay
Glioblastoma
Down Syndrome
Neuroblastoma
Reporter Genes
Genetic Promoter Regions
Intellectual Disability
Embryonic Development
Exons
Carrier Proteins
Transcription Factors

Keywords

  • c-myb
  • Electrophoretic mobility shift assay
  • Genome structure
  • Luciferase

ASJC Scopus subject areas

  • Genetics

Cite this

Molecular mechanisms of human single-minded 2 (SIM2) gene expression : Identification of a promoter site in the SIM2 genomic sequence. / Yamaki, Akiko; Tochigi, Junko; Kudo, Jun; Minoshima, Shinsei; Shimizu, Nobuyoshi; Shimizu, Yoshiko.

In: Gene, Vol. 270, No. 1-2, 30.05.2001, p. 265-275.

Research output: Contribution to journalArticle

Yamaki, Akiko ; Tochigi, Junko ; Kudo, Jun ; Minoshima, Shinsei ; Shimizu, Nobuyoshi ; Shimizu, Yoshiko. / Molecular mechanisms of human single-minded 2 (SIM2) gene expression : Identification of a promoter site in the SIM2 genomic sequence. In: Gene. 2001 ; Vol. 270, No. 1-2. pp. 265-275.
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