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

研究成果: Article

11 引用 (Scopus)

抄録

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.

元の言語English
ページ(範囲)265-275
ページ数11
ジャーナルGene
270
発行部数1-2
DOI
出版物ステータスPublished - 2001 5 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

ASJC Scopus subject areas

  • Genetics

これを引用

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.

:: Gene, 巻 270, 番号 1-2, 30.05.2001, p. 265-275.

研究成果: Article

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. :: Gene. 2001 ; 巻 270, 番号 1-2. pp. 265-275.
@article{c744cf3987dd4782a695384467a01876,
title = "Molecular mechanisms of human single-minded 2 (SIM2) gene expression: Identification of a promoter site in the SIM2 genomic sequence",
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.",
keywords = "c-myb, Electrophoretic mobility shift assay, Genome structure, Luciferase",
author = "Akiko Yamaki and Junko Tochigi and Jun Kudo and Shinsei Minoshima and Nobuyoshi Shimizu and Yoshiko Shimizu",
year = "2001",
month = "5",
day = "30",
doi = "10.1016/S0378-1119(01)00450-4",
language = "English",
volume = "270",
pages = "265--275",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Molecular mechanisms of human single-minded 2 (SIM2) gene expression

T2 - Identification of a promoter site in the SIM2 genomic sequence

AU - Yamaki, Akiko

AU - Tochigi, Junko

AU - Kudo, Jun

AU - Minoshima, Shinsei

AU - Shimizu, Nobuyoshi

AU - Shimizu, Yoshiko

PY - 2001/5/30

Y1 - 2001/5/30

N2 - 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.

AB - 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.

KW - c-myb

KW - Electrophoretic mobility shift assay

KW - Genome structure

KW - Luciferase

UR - http://www.scopus.com/inward/record.url?scp=0035972868&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035972868&partnerID=8YFLogxK

U2 - 10.1016/S0378-1119(01)00450-4

DO - 10.1016/S0378-1119(01)00450-4

M3 - Article

C2 - 11404025

AN - SCOPUS:0035972868

VL - 270

SP - 265

EP - 275

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1-2

ER -