Characterization of the promoter for the human 85 kDa cytosolic phospholipase A2 gene.
The 85 kDa cytosolic phospholipase A2 (cPLA2) plays a key role in the production of arachidonic acid and lysophospholipids, the precursors of eicosanoids and platelet-activating factor. Here we report the cloning of the promoter of the human cPLA2 gene. A 5.7 kb EcoRI fragment containing the most 5' region of the cPLA2 cDNA was sequenced. The transcription initiation site was identified by rapid amplification of 5'-cDNA ends (5'-RACE) and primer extension analysis. DNA sequence analysis of the 595 base pairs 5' of the transcription start site reveals a 48 base purine-pyrimidine dinucleotide repeat (CA repeat), five interferon-gamma response elements (gamma-IRE), one interferon-gamma activated sequence (GAS) and two glucocorticoid response elements (GRE). The promoter lacks a TATA box. It contains a possible CAAT box at -111 and two octamer binding motifs. The 595 base fragment located immediately upstream of the transcriptional start site exhibited functional promoter activity in transient transfection assays in a bronchial epithelial cell line (BEAS 2B cells). Deletion analysis revealed that the CA repeat may confer an inhibitory effect on the cPLA2 promoter activity. The characterization of the human cPLA2 promoter sequence will allow further studies defining the molecular events regulating the expression of the cPLA2 enzyme, especially the cytokine mediated cPLA2 gene expression.
Wu
T
T
Critical Care Medicine Department, National Institutes of Health, Bethesda, MD 20892.
Ikezono
T
T
Angus
C W
CW
Shelhamer
J H
JH
D001483:Base Sequence; D001980:Bronchi; D002460:Cell Line; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000235:genetics*; D054467:Phospholipases A2; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics*; D014162:Transfection
D012333:RNA, Messenger; D010741:Phospholipases A; D054467:Phospholipases A2