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Article

Serum Deprivation-Induced Human GM3 Synthase (hST3Gal V) Gene Expression Is Mediated by Runx2 in Human Osteoblastic MG-63 Cells

by
Hyun-Kyoung Yoon
1,†,
Ji-Won Lee
1,†,
Kyoung-Sook Kim
1,
Seo-Won Mun
1,
Dong-Hyun Kim
1,
Hyun-Jun Kim
2,
Cheorl-Ho Kim
3 and
Young-Choon Lee
1,*
1
Department of Medicinal Biotechnology, College of Health Sciences, Dong-A University, Busan 604-714, Korea
2
Department of Orthopaedic Surgery, College of Medicine, Dong-A University, Busan 604-714, Korea
3
Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, Kyunggi-Do 440-746, Korea
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Int. J. Mol. Sci. 2016, 17(1), 35; https://doi.org/10.3390/ijms17010035
Submission received: 31 October 2015 / Revised: 15 December 2015 / Accepted: 22 December 2015 / Published: 29 December 2015
(This article belongs to the Section Biochemistry)
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Abstract

Serum deprivation (SD) is well known to induce G0/G1 cell cycle arrest and apoptosis in various cells. In the present study, we firstly found that SD could induce G1 arrest and the differentiation of human osteoblastic MG-63 cells, as evidenced by the increase of osteoblastic differentiation markers, such as bone morphogenetic protein-2 (BMP-2), osteocalcin and runt-related transcription factor 2 (Runx2). In parallel, gene expression of human GM3 synthase (hST3Gal V) catalyzing ganglioside GM3 biosynthesis was upregulated by SD in MG-63 cells. The 5′-flanking region of the hST3Gal V gene was functionally characterized to elucidate transcriptional regulation of hST3Gal V in SD-induced MG-63 cells. Promoter analysis using 5′-deletion constructs of the hST3Gal V gene demonstrated that the −432 to −177 region functions as the SD-inducible promoter. Site-directed mutagenesis revealed that the Runx2 binding sites located side-by-side at positions −232 and −222 are essential for the SD-induced expression of hST3Gal V in MG-63 cells. In addition, the chromatin immunoprecipitation assay also showed that Runx2 specifically binds to the hST3Gal V promoter region containing Runx2 binding sites. These results suggest that SD triggers upregulation of hST3Gal V gene expression through Runx2 activation by BMP signaling in MG-63 cells.
Keywords: serum deprivation; human GM3 synthase (hST3Gal V); MG-63 cells; runt-related transcription factor 2 (Runx2); transcriptional regulation serum deprivation; human GM3 synthase (hST3Gal V); MG-63 cells; runt-related transcription factor 2 (Runx2); transcriptional regulation
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MDPI and ACS Style

Yoon, H.-K.; Lee, J.-W.; Kim, K.-S.; Mun, S.-W.; Kim, D.-H.; Kim, H.-J.; Kim, C.-H.; Lee, Y.-C. Serum Deprivation-Induced Human GM3 Synthase (hST3Gal V) Gene Expression Is Mediated by Runx2 in Human Osteoblastic MG-63 Cells. Int. J. Mol. Sci. 2016, 17, 35. https://doi.org/10.3390/ijms17010035

AMA Style

Yoon H-K, Lee J-W, Kim K-S, Mun S-W, Kim D-H, Kim H-J, Kim C-H, Lee Y-C. Serum Deprivation-Induced Human GM3 Synthase (hST3Gal V) Gene Expression Is Mediated by Runx2 in Human Osteoblastic MG-63 Cells. International Journal of Molecular Sciences. 2016; 17(1):35. https://doi.org/10.3390/ijms17010035

Chicago/Turabian Style

Yoon, Hyun-Kyoung, Ji-Won Lee, Kyoung-Sook Kim, Seo-Won Mun, Dong-Hyun Kim, Hyun-Jun Kim, Cheorl-Ho Kim, and Young-Choon Lee. 2016. "Serum Deprivation-Induced Human GM3 Synthase (hST3Gal V) Gene Expression Is Mediated by Runx2 in Human Osteoblastic MG-63 Cells" International Journal of Molecular Sciences 17, no. 1: 35. https://doi.org/10.3390/ijms17010035

APA Style

Yoon, H.-K., Lee, J.-W., Kim, K.-S., Mun, S.-W., Kim, D.-H., Kim, H.-J., Kim, C.-H., & Lee, Y.-C. (2016). Serum Deprivation-Induced Human GM3 Synthase (hST3Gal V) Gene Expression Is Mediated by Runx2 in Human Osteoblastic MG-63 Cells. International Journal of Molecular Sciences, 17(1), 35. https://doi.org/10.3390/ijms17010035

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