Involvement of protein kinase D in phosphorylation and increase of DNA binding of activator protein 2 alpha to downregulate ATP-binding cassette transporter A1.
BACKGROUND
Activator protein (AP) 2alpha negatively regulates expression of ABCA1 gene through Ser-phosphorylation of AP2alpha (Circ Res. 2007;101:156-165). Potential specific Ser-phosphorylation sites for this reaction were investigated in human AP2alpha.


RESULTS
The phosphorylation was shown mediated by PKD, and Ser258 and Ser326 were found in its specific phosphorylation sequence segment in AP2alpha. PKD phosphorylated Ser258 more than Ser326 and induced its binding to the ABCA1 promoter. These reactions and AP2alpha-induced suppression of the ABCA1 promoter activity were reversed by mutation of Ser258 more than Ser326 mutation. Knockdown of PKD by siRNA reduced the AP2alpha 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.


CONCLUSIONS
PKD phosphorylates AP2alpha to negatively regulate expression of ABCA1 gene to increase HDL biogenesis. The major functional phosphorylation of AP2alpha was identified at Ser258 by PKD, in the AP2alpha 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.
Biochemistry, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan.
Noriyuki
N
Iwamoto
Sumiko
S
Abe-Dohmae
Rui
R
Lu
Shinji
S
Yokoyama
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D015536:Down-Regulation; D006801:Humans; D066298:In Vitro Techniques; D008075:Lipoproteins, HDL / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000737:chemistry; D050656:Transcription Factor AP-2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D008075:Lipoproteins, HDL; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D050656:Transcription Factor AP-2; D012694:Serine; D004247:DNA; C089212:protein kinase D; D011493:Protein Kinase C