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Characterization of the 5′ Flanking Region of the Human Complement Factor H Gene
Author(s) -
WILLIAMS S. A.,
VIK D. P.
Publication year - 1997
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1046/j.1365-3083.1997.d01-364.x
Subject(s) - microbiology and biotechnology , biology , 5' flanking region , promoter , gene , enhancer , reporter gene , tata box , caat box , transcription factor , primer extension , transcription (linguistics) , nucleic acid sequence , gene expression , messenger rna , genetics , linguistics , philosophy
The promoter region of the human factor H gene was cloned and a 3 kb Eco RI fragment was sequenced. Primer extension and S1 nuclease analysis were used to determine the transcription start site, which was found to be 10–11 nucleotides upstream of the published cDNA sequence. No canonical TATA or CCAAT boxes were found in conjunction with this site. The sequence from the human H promoter region was compared to that from the mouse gene. There was a region of 800 bp that was 62.5% identical between the two sequences. The sequences of the two promoter regions were compared to a database of transcription factor binding sites. Five elements were identified that matched the consensus sequence 100% and were identical in the two promoter sequences. Promoter assays using the luciferase reporter gene demonstrated that this region contained a functional transcription start site and putative enhancer elements. U118‐MG astroglioma cells and Hep3b hepatoma cells were incubated with various cytokines to measure effects on their factor H mRNA levels. Interferon‐γ (IFN‐γ), but not interleukin‐1 (IL‐1), tumour necrosis factor α (TNF‐α) or IL‐6, was able to increase the level of H mRNA in both cell lines.