title: Hepatocyte nuclear factor 1alpha plays a critical role in PCSK9 gene transcription and regulation by the natural hypocholesterolemic compound berberine.
abstract: PCSK9 is a natural inhibitor of LDL receptor (LDLR) that binds the extracellular domain of LDLR and triggers its intracellular degradation. PCSK9 and LDLR are coordinately regulated at the transcriptional level by sterols through their promoter-imbedded sterol response elements (SRE) and co-induced by statins. Identification of regulatory networks modulating PCSK9 transcription is important for developing selective repressors of PCSK9 to improve statin efficacy by prolonging the up-regulation of LDLR. Interestingly, the plant-derived hypocholesterolemic compound berberine (BBR) up-regulates LDLR expression while down-regulating PCSK9. In our investigations to define mechanisms underlying the transcriptional suppression of PCSK9 by BBR in HepG2 cells, we have identified a highly conserved hepatocyte nuclear factor 1 (HNF1) binding site residing 28 bp upstream from SRE as a critical sequence motif for PCSK9 transcription and its regulation by BBR. Mutation of the HNF1 site reduced PCSK9 promoter activity >90%. A battery of functional assays identified HNF1alpha as the predominant trans-activator for PCSK9 gene working through this sequence motif. We further provide evidence suggesting that HNF1 site works cooperatively with SRE as HNF1 mutation significantly attenuated the activity of nuclear SREBP2 to transactivate PCSK9 promoter. Finally, we show that a coordinate modest reduction of HNF1alpha and nuclear SREBP2 by BBR led to a strong suppression of PCSK9 transcription through these two critical regulatory sequences. This is the first described example of SREBP pairing with HNF1 to control an important regulatory pathway in cholesterol homeostasis. This work also provides a mechanism for how BBR suppresses PCSK9 transcription.
authors.affiliation: Department of Veterans Affairs Palo Alto Health Care System, Palo Alto, California 94304, USA.
authors.forename: Hai
authors.initials: H
authors.lastname: Li
authors.forename: Bin
authors.initials: B
authors.lastname: Dong
authors.forename: Sahng Wook
authors.initials: SW
authors.lastname: Park
authors.forename: Hyun-Sook
authors.initials: HS
authors.lastname: Lee
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Jingwen
authors.initials: J
authors.lastname: Liu
mesh_terms: D000924:Anticholesteremic Agents / Q000494:pharmacology*; D001483:Base Sequence; D001599:Berberine / Q000494:pharmacology*; D001665:Binding Sites; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation; D051538:Hepatocyte Nuclear Factor 1-alpha / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D034741:RNA, Small Interfering / Q000378:metabolism; D011973:Receptors, LDL / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D000924:Anticholesteremic Agents; D051538:Hepatocyte Nuclear Factor 1-alpha; D034741:RNA, Small Interfering; D011973:Receptors, LDL; D001599:Berberine; C472125:PCSK9 protein, human; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D012697:Serine Endopeptidases
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