Premium
Transcriptional regulation of the lysyl oxidase gene by the hypoxia‐inducible factor 1 (HIF1)
Author(s) -
Li Wande,
Zhao Yinzhi,
Gao Song,
Toselli Paul
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.833.16
Subject(s) - lysyl oxidase , microbiology and biotechnology , transcriptional regulation , gene , luciferase , promoter , reporter gene , transcription (linguistics) , chemistry , gene expression , elastin , regulation of gene expression , biology , extracellular matrix , transfection , genetics , biochemistry , linguistics , philosophy
To elucidate transcriptional regulation of lysyl oxidase (LO), a key enzyme for crosslinking collagen and elastin in the extracellular matrix, we cloned the 5'‐flanking region (3,979 nt) of the rat LO gene into the pGL3‐Basic vector upstream of the luciferase gene. 5'‐Deletion assays showed the maximal promoter activity obtained from the 804 base pair fragment upstream of ATG. There are at least 4 hypoxia‐response element (HRE) consensus sequences (ACGTG) existing in this LO promoter region. Treatment of RFL6 lung fibroblasts with cobalt (Co) chloride (100 μM) mimic the hypoxia conditions enhanced LO promoter activities. Dominant negative HIF 1α inhibited LO promoter activities stimulated by Co. Electrophoretic mobility shift, oligonucleotide competition and in vitro translation of HIF 1α assays indicated that only one HRE mapped at −387/−383 relative to the ATG was functionally active among 4 consensuses. Site‐directed mutation of this HRE significantly decreased the LO promoter‐directed expression of the reporter gene. Thus, HIF‐1 activation of the HRE plays a critical role in regulation of LO gene transcription (Supported by NIH grant R01‐ES11340 and Philip Morris ERP).