Characterization of the 5′-flanking fragment of the human GM3-synthase gene

Guichao Zeng, Luoyi Gao, Tian Xia, Tewin Tencomnao, Robert K Yu

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

To investigate the transcriptional regulation of human GM3-synthase, a 5′-flanking fragment of 1379 bp was cloned by a PCR-based procedure. Analysis of the human genomic sequence showed that the gene consists of seven exons, locates at chromosome 2, and spans over 62 kb. There are a number of potential consensus binding sites in the cloned promoter region, but TATA and CCAAT boxes were not found in the promoter. Primer extension analysis identified two transcription start sites approximately 11 and 57 bp upstream of the exon 1. The transcription activity of the promoter was assessed in human HeLa cells by transient transfection. Of the fragments assayed, the proximal 409 bp fragment exhibits the highest transcription activity. Transcription factors that bound to the 409 bp fragment were pulled down by DNA-coupled magnetic beads. Identities of the pull-down proteins were determined by array analysis. Eight transcription factors were identified, which might either bind to the proximal region or be recruited as co-activators of the transcription factor complexes.

Original languageEnglish (US)
Pages (from-to)30-35
Number of pages6
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1625
Issue number1
DOIs
StatePublished - Jan 3 2003

Fingerprint

Transcription Factors
Genes
Transcription
Exons
TATA Box
Chromosomes, Human, Pair 2
Transcription Initiation Site
Chromosomes
HeLa Cells
Genetic Promoter Regions
Transfection
Binding Sites
Polymerase Chain Reaction
DNA
haematoside synthetase
Proteins

Keywords

  • GM3-synthase
  • Ganglioside
  • Promoter
  • Protein/DNA array
  • Sialyltransferase
  • Transcription factor

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology
  • Biophysics

Cite this

Characterization of the 5′-flanking fragment of the human GM3-synthase gene. / Zeng, Guichao; Gao, Luoyi; Xia, Tian; Tencomnao, Tewin; Yu, Robert K.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1625, No. 1, 03.01.2003, p. 30-35.

Research output: Contribution to journalArticle

Zeng, Guichao ; Gao, Luoyi ; Xia, Tian ; Tencomnao, Tewin ; Yu, Robert K. / Characterization of the 5′-flanking fragment of the human GM3-synthase gene. In: Biochimica et Biophysica Acta - Gene Structure and Expression. 2003 ; Vol. 1625, No. 1. pp. 30-35.
@article{24df70d8670f4ab688865e5ebdfac992,
title = "Characterization of the 5′-flanking fragment of the human GM3-synthase gene",
abstract = "To investigate the transcriptional regulation of human GM3-synthase, a 5′-flanking fragment of 1379 bp was cloned by a PCR-based procedure. Analysis of the human genomic sequence showed that the gene consists of seven exons, locates at chromosome 2, and spans over 62 kb. There are a number of potential consensus binding sites in the cloned promoter region, but TATA and CCAAT boxes were not found in the promoter. Primer extension analysis identified two transcription start sites approximately 11 and 57 bp upstream of the exon 1. The transcription activity of the promoter was assessed in human HeLa cells by transient transfection. Of the fragments assayed, the proximal 409 bp fragment exhibits the highest transcription activity. Transcription factors that bound to the 409 bp fragment were pulled down by DNA-coupled magnetic beads. Identities of the pull-down proteins were determined by array analysis. Eight transcription factors were identified, which might either bind to the proximal region or be recruited as co-activators of the transcription factor complexes.",
keywords = "GM3-synthase, Ganglioside, Promoter, Protein/DNA array, Sialyltransferase, Transcription factor",
author = "Guichao Zeng and Luoyi Gao and Tian Xia and Tewin Tencomnao and Yu, {Robert K}",
year = "2003",
month = "1",
day = "3",
doi = "10.1016/S0167-4781(02)00573-0",
language = "English (US)",
volume = "1625",
pages = "30--35",
journal = "Biochimica et Biophysica Acta - Gene Structure and Expression",
issn = "0167-4781",
publisher = "Elsevier BV",
number = "1",

}

TY - JOUR

T1 - Characterization of the 5′-flanking fragment of the human GM3-synthase gene

AU - Zeng, Guichao

AU - Gao, Luoyi

AU - Xia, Tian

AU - Tencomnao, Tewin

AU - Yu, Robert K

PY - 2003/1/3

Y1 - 2003/1/3

N2 - To investigate the transcriptional regulation of human GM3-synthase, a 5′-flanking fragment of 1379 bp was cloned by a PCR-based procedure. Analysis of the human genomic sequence showed that the gene consists of seven exons, locates at chromosome 2, and spans over 62 kb. There are a number of potential consensus binding sites in the cloned promoter region, but TATA and CCAAT boxes were not found in the promoter. Primer extension analysis identified two transcription start sites approximately 11 and 57 bp upstream of the exon 1. The transcription activity of the promoter was assessed in human HeLa cells by transient transfection. Of the fragments assayed, the proximal 409 bp fragment exhibits the highest transcription activity. Transcription factors that bound to the 409 bp fragment were pulled down by DNA-coupled magnetic beads. Identities of the pull-down proteins were determined by array analysis. Eight transcription factors were identified, which might either bind to the proximal region or be recruited as co-activators of the transcription factor complexes.

AB - To investigate the transcriptional regulation of human GM3-synthase, a 5′-flanking fragment of 1379 bp was cloned by a PCR-based procedure. Analysis of the human genomic sequence showed that the gene consists of seven exons, locates at chromosome 2, and spans over 62 kb. There are a number of potential consensus binding sites in the cloned promoter region, but TATA and CCAAT boxes were not found in the promoter. Primer extension analysis identified two transcription start sites approximately 11 and 57 bp upstream of the exon 1. The transcription activity of the promoter was assessed in human HeLa cells by transient transfection. Of the fragments assayed, the proximal 409 bp fragment exhibits the highest transcription activity. Transcription factors that bound to the 409 bp fragment were pulled down by DNA-coupled magnetic beads. Identities of the pull-down proteins were determined by array analysis. Eight transcription factors were identified, which might either bind to the proximal region or be recruited as co-activators of the transcription factor complexes.

KW - GM3-synthase

KW - Ganglioside

KW - Promoter

KW - Protein/DNA array

KW - Sialyltransferase

KW - Transcription factor

UR - http://www.scopus.com/inward/record.url?scp=0037415120&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037415120&partnerID=8YFLogxK

U2 - 10.1016/S0167-4781(02)00573-0

DO - 10.1016/S0167-4781(02)00573-0

M3 - Article

VL - 1625

SP - 30

EP - 35

JO - Biochimica et Biophysica Acta - Gene Structure and Expression

JF - Biochimica et Biophysica Acta - Gene Structure and Expression

SN - 0167-4781

IS - 1

ER -