Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter

Nobuyuki Shiomi, Fumio Myokai, Koji Naruishi, Kosuke Oyaizu, Kyoko Senoo, Tomoko Yamaguchi, Salomon Amar, Shogo Takashiba

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor α factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-α expression in the human monocytic cell line, THP-1. To characterize the human (h) LITAF promoter, we isolated a 1.2-kb DNA fragment and followed this by a screening of human genomic DNA with a hLITAF cDNA probe. A 34-bp sequence domain located from nucleotides -74 to -43 in the hLITAF promoter exhibited the highest basal reporter gene activity; however, the activity was not elevated by lipopolysaccharide (LPS) stimulation. The sequence domain included a consensus sequence for hepatocyte nuclear factor (HNF)-3α, regulating the transcription of many kinds of genes. Interestingly, the DNA sequence position between -542 and -538 in the hLITAF promoter contained the CTCCC motif, which has been reported to act as a specific binding site for hLITAF protein. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of THP-1 nuclear factors to a 22 bp probe containing the CTCCC motif. In addition, hLITAF mRNA and nuclear hLITAF protein increased significantly in the THP-1 cells immediately after LPS stimulation. These results suggest that the consensus sequence for HNF-3α, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.

Original languageEnglish
Pages (from-to)360-368
Number of pages9
JournalFEMS Immunology and Medical Microbiology
Volume47
Issue number3
DOIs
Publication statusPublished - Aug 2006

Fingerprint

Lipopolysaccharides
Organism Cloning
Tumor Necrosis Factor-alpha
Hepatocyte Nuclear Factors
Consensus Sequence
Nuclear Proteins
NFI Transcription Factors
DNA
Electrophoretic Mobility Shift Assay
Reporter Genes
Carrier Proteins
Transcription Factors
Necrosis
Nucleotides
Complementary DNA
Binding Sites
Cell Line
Messenger RNA
Genes
Neoplasms

Keywords

  • Electrophoretic mobility shift assay
  • Genomic cloning
  • LPS-induced TNF-α factor
  • Promoter sequence
  • Reporter gene assay

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Infectious Diseases

Cite this

Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter. / Shiomi, Nobuyuki; Myokai, Fumio; Naruishi, Koji; Oyaizu, Kosuke; Senoo, Kyoko; Yamaguchi, Tomoko; Amar, Salomon; Takashiba, Shogo.

In: FEMS Immunology and Medical Microbiology, Vol. 47, No. 3, 08.2006, p. 360-368.

Research output: Contribution to journalArticle

Shiomi, Nobuyuki ; Myokai, Fumio ; Naruishi, Koji ; Oyaizu, Kosuke ; Senoo, Kyoko ; Yamaguchi, Tomoko ; Amar, Salomon ; Takashiba, Shogo. / Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter. In: FEMS Immunology and Medical Microbiology. 2006 ; Vol. 47, No. 3. pp. 360-368.
@article{1d761c78ec8b4fd28136bce4034e4f08,
title = "Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter",
abstract = "We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor α factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-α expression in the human monocytic cell line, THP-1. To characterize the human (h) LITAF promoter, we isolated a 1.2-kb DNA fragment and followed this by a screening of human genomic DNA with a hLITAF cDNA probe. A 34-bp sequence domain located from nucleotides -74 to -43 in the hLITAF promoter exhibited the highest basal reporter gene activity; however, the activity was not elevated by lipopolysaccharide (LPS) stimulation. The sequence domain included a consensus sequence for hepatocyte nuclear factor (HNF)-3α, regulating the transcription of many kinds of genes. Interestingly, the DNA sequence position between -542 and -538 in the hLITAF promoter contained the CTCCC motif, which has been reported to act as a specific binding site for hLITAF protein. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of THP-1 nuclear factors to a 22 bp probe containing the CTCCC motif. In addition, hLITAF mRNA and nuclear hLITAF protein increased significantly in the THP-1 cells immediately after LPS stimulation. These results suggest that the consensus sequence for HNF-3α, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.",
keywords = "Electrophoretic mobility shift assay, Genomic cloning, LPS-induced TNF-α factor, Promoter sequence, Reporter gene assay",
author = "Nobuyuki Shiomi and Fumio Myokai and Koji Naruishi and Kosuke Oyaizu and Kyoko Senoo and Tomoko Yamaguchi and Salomon Amar and Shogo Takashiba",
year = "2006",
month = "8",
doi = "10.1111/j.1574-695X.2006.00094.x",
language = "English",
volume = "47",
pages = "360--368",
journal = "Pathogens and Disease",
issn = "2049-632X",
publisher = "John Wiley & Sons Inc.",
number = "3",

}

TY - JOUR

T1 - Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter

AU - Shiomi, Nobuyuki

AU - Myokai, Fumio

AU - Naruishi, Koji

AU - Oyaizu, Kosuke

AU - Senoo, Kyoko

AU - Yamaguchi, Tomoko

AU - Amar, Salomon

AU - Takashiba, Shogo

PY - 2006/8

Y1 - 2006/8

N2 - We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor α factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-α expression in the human monocytic cell line, THP-1. To characterize the human (h) LITAF promoter, we isolated a 1.2-kb DNA fragment and followed this by a screening of human genomic DNA with a hLITAF cDNA probe. A 34-bp sequence domain located from nucleotides -74 to -43 in the hLITAF promoter exhibited the highest basal reporter gene activity; however, the activity was not elevated by lipopolysaccharide (LPS) stimulation. The sequence domain included a consensus sequence for hepatocyte nuclear factor (HNF)-3α, regulating the transcription of many kinds of genes. Interestingly, the DNA sequence position between -542 and -538 in the hLITAF promoter contained the CTCCC motif, which has been reported to act as a specific binding site for hLITAF protein. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of THP-1 nuclear factors to a 22 bp probe containing the CTCCC motif. In addition, hLITAF mRNA and nuclear hLITAF protein increased significantly in the THP-1 cells immediately after LPS stimulation. These results suggest that the consensus sequence for HNF-3α, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.

AB - We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor α factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-α expression in the human monocytic cell line, THP-1. To characterize the human (h) LITAF promoter, we isolated a 1.2-kb DNA fragment and followed this by a screening of human genomic DNA with a hLITAF cDNA probe. A 34-bp sequence domain located from nucleotides -74 to -43 in the hLITAF promoter exhibited the highest basal reporter gene activity; however, the activity was not elevated by lipopolysaccharide (LPS) stimulation. The sequence domain included a consensus sequence for hepatocyte nuclear factor (HNF)-3α, regulating the transcription of many kinds of genes. Interestingly, the DNA sequence position between -542 and -538 in the hLITAF promoter contained the CTCCC motif, which has been reported to act as a specific binding site for hLITAF protein. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of THP-1 nuclear factors to a 22 bp probe containing the CTCCC motif. In addition, hLITAF mRNA and nuclear hLITAF protein increased significantly in the THP-1 cells immediately after LPS stimulation. These results suggest that the consensus sequence for HNF-3α, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.

KW - Electrophoretic mobility shift assay

KW - Genomic cloning

KW - LPS-induced TNF-α factor

KW - Promoter sequence

KW - Reporter gene assay

UR - http://www.scopus.com/inward/record.url?scp=33745811705&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745811705&partnerID=8YFLogxK

U2 - 10.1111/j.1574-695X.2006.00094.x

DO - 10.1111/j.1574-695X.2006.00094.x

M3 - Article

C2 - 16872372

AN - SCOPUS:33745811705

VL - 47

SP - 360

EP - 368

JO - Pathogens and Disease

JF - Pathogens and Disease

SN - 2049-632X

IS - 3

ER -