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Housekeeping promoter 5’pcmah-2 of pig CMP-N-acetylneuraminic acid hydroxylase gene for NeuGc expression

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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.

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Acknowledgments

This study has been in part supported from the Basic Science Research Program through National Research Foundation of Korea (NRF) grant (NRF-2015R1D1A1A01057153).

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Authors and Affiliations

  1. Molecular and Cellular Glycobiology Unit, Department of Biological Science, Sungkyunkwan University, Chunchun-Dong, Suwon, 16419, South Korea

    Kwon-Ho Song, Choong-Hwan Kwak, Un-Ho Jin, Sun-Hyung Ha, Jun-Young Park, Fukushi Abekura & Cheorl-Ho Kim

  2. Research Institute of Biomedical Engineering and Department of Medicine, Catholic University of Daegu School of Medicine, Daegu, 705-034, South Korea

    Young-Chae Chang

  3. Department of Enteric Bacterial Infections, Korea Centers for Disease Control and Prevention, Cheongwon-gun, South Korea

    Seung-Hak Cho

  4. Functional Genomics Laboratory, Department of Animal Sciences, The Ohio State University, 2029 Fyffe Court, Columbus, OH, 43210, USA

    Kichoon Lee

  5. Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, South Korea

    Tae-Wook Chung & Ki-Tae Ha

  6. Faculty of Medicinal Biotechnology, Dong-A University, Busan, 604-714, South Korea

    Young-Choon Lee

  7. Department of Medical Device Management and Research, Samsung Advanced Institute for Health Sciences & Technology (SAIHST), Sungkyunkwan University, Seoul, 06351, South Korea

    Cheorl-Ho Kim

  8. Department of Biological Science, Sungkyunkwan University, 2066, Seobu-Ro, Suwon, Gyunggi-Do, 16419, South Korea

    Cheorl-Ho Kim

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  1. Kwon-Ho Song
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  2. Choong-Hwan Kwak
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  6. Fukushi Abekura
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  9. Kichoon Lee
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  10. Tae-Wook Chung
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  11. Ki-Tae Ha
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  13. Cheorl-Ho Kim
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Correspondence to Cheorl-Ho Kim.

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Kwon-Ho Song and Choong-Hwan Kwak contributed equally as the co-1st authors.

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Song, KH., Kwak, CH., Jin, UH. et al. Housekeeping promoter 5’pcmah-2 of pig CMP-N-acetylneuraminic acid hydroxylase gene for NeuGc expression. Glycoconj J 33, 779–788 (2016). https://doi.org/10.1007/s10719-016-9671-5

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  • DOI: https://doi.org/10.1007/s10719-016-9671-5

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