title: Housekeeping promoter 5'pcmah-2 of pig CMP-N-acetylneuraminic acid hydroxylase gene for NeuGc expression.
abstract: In the present study, we isolated pCMAH house-keeping promoter regions (Ph), which are responsible for transcriptional regulation and which are located upstream of the alternative transcript pcmah-2. Luciferase reporter assays using serial construction of each deleted promoter demonstrated that the Ph promoter was highly active in pig-derived kidney PK15. Ph promoter of pcmah lacked a TATA box, but contained three putative Sp1 binding sites. Mutations of these Sp1 binding sites always resulted in the reduction of luciferase activities in Ph-334. In addition, treatment with mithramycin A (25-100 nM) decreased the luciferase activities of the Ph promoters and NeuGc expression in a dose-dependent manner. Electrophoretic mobility shift assay analysis revealed that the probes containing each Sp1 binding site bound to Sp1. Taken together, the results indicate that Sp1 bind to their putative binding sites on the Ph promoter regions of pcmah and positively regulate the promoter activity in pig kidney cells. Interspecies comparison of 5'UTRs and 5'flanking regions shows high homology between pig and cattle, and Sp1 binding sites existing in genomic regions corresponding Ph region are evolutionally conserved.
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Kwon-Ho
authors.initials: KH
authors.lastname: Song
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Choong-Hwan
authors.initials: CH
authors.lastname: Kwak
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Un-Ho
authors.initials: UH
authors.lastname: Jin
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Sun-Hyung
authors.initials: SH
authors.lastname: Ha
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea.
authors.forename: Fukushi
authors.initials: F
authors.lastname: Abekura
authors.affiliation: Research Institute of Biomedical Engineering and Department of Medicine, Catholic University of Daegu School of Medicine, Daegu, 705-034, South Korea.
authors.forename: Young-Chae
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Department of Enteric Bacterial Infections, Korea Centers for Disease Control and Prevention, Cheongwon-gun, South Korea.
authors.forename: Seung-Hak
authors.initials: SH
authors.lastname: Cho
authors.affiliation: Functional Genomics Laboratory, Department of Animal Sciences, The Ohio State University, 2029 Fyffe Court, Columbus, OH, 43210, USA.
authors.forename: Kichoon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, South Korea.
authors.forename: Tae-Wook
authors.initials: TW
authors.lastname: Chung
authors.affiliation: Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, South Korea.
authors.forename: Ki-Tae
authors.initials: KT
authors.lastname: Ha
authors.affiliation: Faculty of Medicinal Biotechnology, Dong-A University, Busan, 604-714, South Korea.
authors.forename: Young-Choon
authors.initials: YC
authors.lastname: Lee
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea. chkimbio@skku.edu.|Department of Medical Device Management and Research, Samsung Advanced Institute for Health Sciences & Technology (SAIHST), Sungkyunkwan University, Seoul, 06351, South Korea. chkimbio@skku.edu.|Department of Biological Science, Sungkyunkwan University, 2066, Seobu-Ro, Suwon, Gyunggi-Do, 16419, South Korea. chkimbio@skku.edu.
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D015971:Gene Expression Regulation, Enzymologic; D020043:Genes, Essential / Q000502:physiology*; D006899:Mixed Function Oxygenases / Q000096:biosynthesis*; D009438:Neuraminic Acids / Q000378:metabolism*; D008926:Plicamycin / Q000494:pharmacology; D020218:Response Elements / Q000502:physiology*; D013552:Swine
chemical_list: D009438:Neuraminic Acids; C032592:N-glycolylneuraminic acid; D006899:Mixed Function Oxygenases; C074383:CMPacetylneuraminate monooxygenase; D008926:Plicamycin
keywords: CMP-N-acetylneuraminic acid hydroxylase (CMAH); N-glycolylneuraminic acid (NeuGc); Specific protein 1 (Sp1)