on October 9, 2008
Published online before print October 9, 2008, doi: 10.1161/ATVBAHA.108.174714
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Submitted on July 25, 2008
Accepted on September 29, 2008
Involvement of Protein Kinase D in Phosphorylation and Increase of DNA Binding of Activator Protein 2
to Downregulate ATP-Binding Cassette Transporter A1
Noriyuki Iwamoto ;
From Biochemistry, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
* To whom correspondence should be addressed. E-mail: syokoyam{at}med.nagoya-cu.ac.jp.
Background—Activator protein (AP) 2
negatively regulates expression of ABCA1 gene through Ser-phosphorylation of AP2 (Circ Res. 2007;101:156–165). Potential specific Ser-phosphorylation sites for this reaction were investigated in human AP2.
Methods and Results—The phosphorylation was shown mediated by PKD, and Ser258 and Ser326 were found in its specific phosphorylation sequence segment in AP2. PKD phosphorylated Ser258 more than Ser326 and induced its binding to the ABCA1 promoter. These reactions and AP2-induced suppression of the ABCA1 promoter activity were reversed by mutation of Ser258 more than Ser326 mutation. Knockdown of PKD by siRNA reduced the AP2 Ser-phosphorylation, and increased ABCA1 expression and HDL biogenesis. Gö6983 inhibited PKD more selectively than PKC in THP-1 and HEK 293 cells and in mice, and increased ABCA1 expression, HDL biogenesis, and plasma HDL level.
Conclusion—PKD phosphorylates AP2 to negatively regulate expression of ABCA1 gene to increase HDL biogenesis. The major functional phosphorylation of AP2 was identified at Ser258 by PKD, in the AP2 basic domain highly conserved among species and all 5 subtypes of AP2. PKD/AP2 system can be a potent pharmacological target for prevention of atherosclerosis.
Key words: HDL • ABCA1 • AP2
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PKD •
doxazosin •
atherosclerosis