The role of TFIID, the initiator element and a novel 5' TFIID binding site in the transcriptional control of the TATA-less human cytosolic phospholipase A2-alpha promoter.
Human cytosolic phospholipase A2-alpha (cPLA2-alpha) is a critical enzyme in the liberation of arachidonic acid (AA) from cellular membranes and the subsequent formation of prostaglandins (PGs), leukotrienes (LTs), hydroxyeicosatetraenoic acids (HETEs) and platelet activating factor in many different cell types. Much is known of the effect of posttranslational phosphorylation and calcium binding events on the enzymatic activity of cPLA2-alpha, but to date little is known about its specific transcriptional control. Through the use of reporter gene constructs and eletrophoretic mobility shift assays (EMSAs), this study determined the minimal promoter required for basal transcriptional activity of the human cPLA2-alpha promoter to include base pairs -40 through the transcription start site (TSS). In addition, it confirms the importance of an initiator (Inr) element at the TSS by deletion reporter gene analysis, and further identifies bases -3 (C) and -2 (T) as critical bases in the Inr function by mutation reporter gene analysis. Finally, this study describes a novel AAGGAG motif at -30 to -35 which is bound by TATA-box binding protein (TBP) and is critical for basal transcriptional activity.
Cowan
Mark J
MJ
Division of Pulmonary and Critical Care Medicine, The University of Maryland, 10 North Greene Street, Room 3D-127, Baltimore, MD 21201, USA. mark.cowan@verizon.net
Yao
Xiang-Lan
XL
Pawliczak
Rafal
R
Huang
Xiuli
X
Logun
Carolea
C
Madara
Patricia
P
Alsaaty
Sura
S
Wu
Tong
T
Shelhamer
James H
JH
D001665:Binding Sites; D001980:Bronchi / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D024202:Electrophoretic Mobility Shift Assay; D004848:Epithelium / Q000378:metabolism; D005809:Genes, Regulator / Q000235:genetics*; D054513:Group IV Phospholipases A2; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D009154:Mutation; D010741:Phospholipases A / Q000235:genetics*; D054467:Phospholipases A2; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017384:Sequence Deletion; D016385:TATA Box; D035362:Transcription Factor TFIID / Q000235:genetics / Q000378:metabolism*; D024363:Transcription Initiation Site / Q000502:physiology*; D014158:Transcription, Genetic
D035362:Transcription Factor TFIID; D008156:Luciferases; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2