title: HTRA1 (high temperature requirement A serine peptidase 1) gene is transcriptionally regulated by insertion/deletion nucleotides located at the 3' end of the ARMS2 (age-related maculopathy susceptibility 2) gene in patients with age-related macular degeneration.
abstract: Dry age-related macular degeneration (AMD) accounts for over 85% of AMD cases in the United States, whereas Japanese AMD patients predominantly progress to wet AMD or polypoidal choroidal vasculopathy. Recent genome-wide association studies have revealed a strong association between AMD and an insertion/deletion sequence between the ARMS2 (age-related maculopathy susceptibility 2) and HTRA1 (high temperature requirement A serine peptidase 1) genes. Transcription regulator activity was localized in mouse retinas using heterozygous HtrA1 knock-out mice in which HtrA1 exon 1 was replaced with β-galactosidase cDNA, thereby resulting in dominant expression of the photoreceptors. The insertion/deletion sequence significantly induced HTRA1 transcription regulator activity in photoreceptor cell lines but not in retinal pigmented epithelium or other cell types. A deletion construct of the HTRA1 regulatory region indicated that potential transcriptional suppressors and activators surround the insertion/deletion sequence. Ten double-stranded DNA probes for this region were designed, three of which interacted with nuclear extracts from 661W cells in EMSA. Liquid chromatography-mass spectrometry (LC-MS/MS) of these EMSA bands subsequently identified a protein that bound the insertion/deletion sequence, LYRIC (lysine-rich CEACAM1 co-isolated) protein. In addition, induced pluripotent stem cells from wet AMD patients carrying the insertion/deletion sequence showed significant up-regulation of the HTRA1 transcript compared with controls. These data suggest that the insertion/deletion sequence alters the suppressor and activator cis-elements of HTRA1 and triggers sustained up-regulation of HTRA1. These results are consistent with a transgenic mouse model that ubiquitously overexpresses HtrA1 and exhibits characteristics similar to those of wet AMD patients.
authors.affiliation: From the Division of Molecular and Cellular Biology, National Institute of Sensory Organs, and.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Iejima
authors.affiliation: From the Division of Molecular and Cellular Biology, National Institute of Sensory Organs, and.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Itabashi
authors.affiliation: From the Division of Molecular and Cellular Biology, National Institute of Sensory Organs, and.
authors.forename: Yuich
authors.initials: Y
authors.lastname: Kawamura
authors.affiliation: the Division of Ophthalmology, National Hospital Organization Tokyo Medical Center, Tokyo 152-8902, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Noda
authors.affiliation: the Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan, and.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Yuasa
authors.affiliation: the Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan, and.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fukuda
authors.affiliation: the Division of Gene Function in Animals, Nara Institute of Science and Technology, Nara 630-0101, Japan.
authors.forename: Chio
authors.initials: C
authors.lastname: Oka
authors.affiliation: From the Division of Molecular and Cellular Biology, National Institute of Sensory Organs, and takeshi_iwata@kankakuki.go.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwata
mesh_terms: D000368:Aged; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002853:Chromatography, Liquid; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D000074289:High-Temperature Requirement A Serine Peptidase 1; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012160:Retina / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: C514392:ARMS2 protein, human; D011506:Proteins; D000074289:High-Temperature Requirement A Serine Peptidase 1; C474012:HTRA1 protein, human; D012697:Serine Endopeptidases
keywords: ARMS2; Age-related Macular Degeneration; DNA-binding Protein; EMSA; HTRA1; Photoreceptor; Retina; Retinal Degeneration; Transcription Regulation