title: Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene.
abstract: We previously cloned a molecule that interacts with angiotensin II type 1 (AT1) receptor to exert an inhibitory function on AT1 receptor signaling that we named ATRAP/Agtrap (for AT1 receptor-associated protein). In the present study we examined the regulation of basal ATRAP gene expression using renal distal convoluted tubule cells. We found that serum starvation upregulated basal expression of ATRAP gene, a response that required de novo mRNA and protein synthesis. Luciferase assay revealed that the proximal promoter region directs transcription and that a putative binding site of runt-related transcription factors (RBE) is important for transcriptional activation. The results of RBE-decoy transfection and endogenous knockdown by small interference RNA showed that the runt-related transcription factor Runx3 is involved in ATRAP gene expression. Chromatin immunoprecipitation assay also supported the binding of Runx3 to the ATRAP promoter in renal distal convoluted tubule cells. Immunohistochemistry demonstrated the expression of Runx3 and ATRAP proteins in the distal convoluted and connecting tubules of the kidney in consecutive sections. Furthermore, the Runx3 immunostaining was decreased together with a concomitant suppression of ATRAP expression in the affected kidney after 7 days of unilateral ureteral obstruction. These findings indicate that Runx3 plays a role in ATRAP gene expression in renal distal tubular cells both in vitro and in vivo.
authors.affiliation: Department of Medical Science and Cardiorenal Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Miyuki
authors.initials: M
authors.lastname: Matsuda
authors.forename: Kouichi
authors.initials: K
authors.lastname: Tamura
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Wakui
authors.forename: Toru
authors.initials: T
authors.lastname: Dejima
authors.forename: Akinobu
authors.initials: A
authors.lastname: Maeda
authors.forename: Masato
authors.initials: M
authors.lastname: Ohsawa
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Kanaoka
authors.forename: Sona
authors.initials: S
authors.lastname: Haku
authors.forename: Kengo
authors.initials: K
authors.lastname: Azushima
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamasaki
authors.forename: Daisuke
authors.initials: D
authors.lastname: Saito
authors.forename: Tomonori
authors.initials: T
authors.lastname: Hirose
authors.forename: Yohei
authors.initials: Y
authors.lastname: Maeshima
authors.forename: Yoji
authors.initials: Y
authors.lastname: Nagashima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Umemura
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D050677:Core Binding Factor Alpha 3 Subunit / Q000378:metabolism*; D016895:Culture Media, Serum-Free; D055785:Gene Knockdown Techniques; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051785:Smad Proteins / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000235:genetics*; D015854:Up-Regulation / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C119712:Agtrap protein, mouse; D050677:Core Binding Factor Alpha 3 Subunit; D016895:Culture Media, Serum-Free; D012333:RNA, Messenger; C493730:Runx3 protein, mouse; D051785:Smad Proteins