title: Transcriptional regulation of podoplanin expression by Prox1 in lymphatic endothelial cells.
abstract: Transcription factor prospero homeobox 1 (Prox-1) and podoplanin (PDPN), mucin-type transmembane protein, are both constantly expressed in lymphatic endothelial cells (LECs) and appear to function in an LEC-autonomous manner. Mice globally lacking PDPN (Pdpn(-/-)) develop abnormal and blood-filled lymphatic vessels that highly resemble those in inducible mice lacking Prox-1 (Prox1(-/-)). Prox1 has also been reported to induce PDPN expression in cultured ECs. Thus, we hypothesize that PDPN functions downstream of Prox1 and that its expression is regulated by Prox1 in LECs at the transcriptional level. We first identified four putative binding elements for Prox1 in the 5' upstream regulatory region of Pdpn gene and found that Prox1 directly binds to the 5' regulatory sequence of Pdpn gene in LECs by chromatin immunoprecipitation assay. DNA pull down assay confirmed that Prox1 binds to the putative binding element. In addition, luciferase reporter assay indicated that Prox1 binding to the 5' regulatory sequence of Pdpn regulates Pdpn gene expression. We are therefore the first to experimentally demonstrate that Prox1 regulates PDPN expression at the transcriptional level in the lymphatic vascular system.
authors.affiliation: Department of Non-Communicable Diseases Control and Prevention, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou City, Zhejiang, P.R. China. Electronic address: yfpan@cdc.zj.cn.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Pediatrics, Qingdao Women's & Children's Hospital, Qingdao City, Shandong, P.R.China.
authors.forename: Wen-di
authors.initials: WD
authors.lastname: Wang
authors.affiliation: Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA. Electronic address: Tadayuki-Yago@omrf.org.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yago
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D047369:Chromatin Immunoprecipitation; D042783:Endothelial Cells / Q000166:cytology*; D015870:Gene Expression; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D000098551:Prospero-Related Homeobox 1 Protein; D000098853:Podoplanin
chemical_list: D020121:5' Untranslated Regions; D018398:Homeodomain Proteins; D008562:Membrane Glycoproteins; D025521:Tumor Suppressor Proteins; D000098551:Prospero-Related Homeobox 1 Protein; C490441:Pdpn protein, mouse; D000098853:Podoplanin; C117468:PDPN protein, human; C092025:Pdpn protein, rat
keywords: DNA pull down assay; HEK 293T; Prox1; chromatin immunoprecipitation; luciferase reporter assay; lymphatic endothelial cells; lymphatic vascular system; mutagenesis; podoplanin; transcriptional regulation