Prohibitins, novel vitamin K2 target factors in osteoblast

Tatsuya Uebi, Makoto Umeda, Naoya Maekawa, Satoshi Karasawa, Hiroshi Handa, Takeshi Imai

Department of Aging Intervention, National Center for Geriatrics and Gerontology (NCGG), Obu, Japan.
Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.

DOI: 10.4236/jbm.2013.13001                

Abstract

Vitamin K2 (VK2, menaquinone) is a drug for osteoporosis. VK2 acts as a cofactor for γ-glutamyl carboxylase, which catalyzes the carboxylation of specific glutamic acid residues (γ-carboxylation) of substrate proteins. Here we demonstrate that VK2 also regulate osteoblastgenic marker gene expression. Using VK2-immobilzed nanobeads new target proteins were purified and identified from osteoblastic cell line. They are prohibitin 1 and 2 (PHB1 & 2), respectively. To confirm the PHBs function on VK2-dependent transcription, PHB1 & 2 were knock-down and osteocalcin gene 2 transcriptions were analyzed, indicating that PHBs regulate VK2-dependent transcription. Taken together PHBs are VK2 target proteins for osteoblastgenic transcription.

Keywords

Vitamin K2; Prohibitin; Osteoblast; Runt-Related Transcription Factor 2 (Runx2)

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Dam, H. (1929) Cholesterinstoffwechsel in huhnereiern und h?hnchen. Biochemische Zeitschrift, 215, 475-492.
[2]	Karasawa, S., Azuma, M., Kasama, T., Sakamoto, S., Kabe, Y., Imai, T., Yamaguchi, Y., Miyazawa, K. and Handa, H. (2013) Vitamin K2 covalently binds to Bak and induces Bak-mediated apoptosis. Molecular Pharmaceutics, 83, 613-620. http://dx.doi.org/10.1124/mol.112.082602
[3]	Seegers, W.H. and Bang, N.U. (1967) Blood clotting enzymology. Academic Press, New York.
[4]	Dam, H. (1935) The antihaemorrhagic vitamin of the chick. Biochemical Journal, 29, 1273-1285.
[5]	Hauschka, P.V., Lian, J.B. and Gallop, P.M. (1975) Direct identification of the calciumbinding amino acid, gamma-carboxyglutamate, in mineralized tissue. Proceedings of the National Academy of Sciences of the United States of America, 72, 3925-3929. http://dx.doi.org/10.1073/pnas.72.10.3925
[6]	Thorp, J.A., Gaston, L., Caspers, D.R. and Pal, M.L. (1995) Current concepts and controversies in the use of vitamin K. Drugs, 49, 376-387. http://dx.doi.org/10.2165/00003495-199549030-00005
[7]	Iwamoto, J., Takeda, T. and Sato, Y. (2004) Effects of vitamin K2 on osteoporosis. Current Pharmaceutical De- sign, 10, 2557-2576. http://dx.doi.org/10.2174/1381612043383782
[8]	Ushiroyama, T., Ikeda, A. and Ueki, M. (2002) Effect of continuous combined therapy with vitamin K(2) and vitamin D(3) on bone mineral density and coagulofibrinolysis function in postmenopausal women. Maturitas, 41, 211-221. http://dx.doi.org/10.1016/S0378-5122(01)00275-4
[9]	Sudo, H., Kodama, H., Amagai, Y., Yamamoto, S. and Kasai, S. (1983). In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. The Journal of Cell Biology, 96, 191- 198. http://dx.doi.org/10.1083/jcb.96.1.191
[10]	Takai, H., Kanematsu, M., Yano, K., Tsuda, E., Higashio, K., Ikeda, K., Watanabe, K. and Yamada, Y. (1998) Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastogenesis inhibitory factor by bone marrow stromal cells. The Journal of Biological Chemistry, 273, 27091-27096. http://dx.doi.org/10.1074/jbc.273.42.27091
[11]	Jochum, W., David, J.P., Elliott, C., Wutz, A., Plenk Jr., H., Matsuo, K. and Wagner, E.F. (2000). Increased bone formation and osteosclerosis in mice overexpressing the transcription factor Fra-1. Nature Medicine, 6, 980-984. http://dx.doi.org/10.1038/79676
[12]	Dignam, J.D., Lebovitz, R.M. and Roeder, R.G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Research, 11, 1475-1489. http://dx.doi.org/10.1093/nar/11.5.1475
[13]	Maekawa, N., Hiramoto, M., Sakamoto, S., Azuma, M., Ito, T., Ikeda, M., Naitou, M., Acharya, H.P., Kobayashi, Y., Suematsu, M., Handa, H. and Imai, T. (2011) High-performance affinity purification for identification of 15-deoxy-Δ12,14-PGJ2 interacting factors using magnetic nanobeads. Biomedical Chromatography, 25, 466-471. http://dx.doi.org/10.1093/nar/11.5.1475
[14]	Ducy, P. and Karsenty, G. (1995) Two distinct osteob- last-specific cisacting elements control expression of a mouse osteocalcin gene. Molecular and Cellular Biology, 15, 1858-1869.
[15]	Geoffroy, V., Ducy, P. and Karsenty, G. (1995) A PEBP2a/AML-1-related factor increases osteocalcin promoter activity through its binding to an osteoblast-specific cisacting element. The Journal of Biological Chemistry, 270, 30973-30979. http://dx.doi.org/10.1074/jbc.270.52.30973
[16]	Ducy, P., Zhang, R., Geoffroy, V., Ridall, A.L. and Karsenty, G. (1997) Osf2/Cbfa1: A transcriptional activator of osteoblast differentiation. Cell, 89, 747-754. http://dx.doi.org/10.1016/S0092-8674(00)80257-3
[17]	Komori, T., Yagi, H., Nomura, S., Yamaguchi, A., Sasaki, K., Deguchi, K., Shimizu, Y., Bronson, R.T., Gao, Y.H., Inada, M., Sato, M., Okamoto, R., Kitamura, Y., Yoshiki, S. and Kishimoto, T. (1997). Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell, 89, 755-764. http://dx.doi.org/10.1016/S0092-8674(00)80258-5
[18]	Hiramoto, M., Maekawa, N., Kuge, T., Ayabe, F., Watanabe, A., Masaike, Y., Hatakeyama, M., Handa, H. and Imai, T. (2010) High-performance affinity chromatography method for identification of L-arginine interacting factors using magnetic nanobeads. Biomedical Chromatography, 24, 606-612. http://dx.doi.org/10.1002/bmc.1334
[19]	Umeda, M., Uebi, T., Maekawa, N., Masaike, Y., Handa, H. and Imai, T. (2013) Effective cofactor complex purification using nanobeads. Journal of Bioscience and Medicine, in Press.
[20]	Otto, F., Thornell, A.P., Crompton, T., Denzel, A., Gilmour, K.C., Rosewell, I.R., Stamp, G.W.H., Beddington, R.S.P., Mundlos, S., Olsen, B.R., Selby, P.B. and Owen, M.J. (1997). Cbfa1, a Candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell, 89, 765-771. http://dx.doi.org/10.1016/S0092-8674(00)80259-7
[21]	He, B., Feng, Q., Mukherjee, A., Lonard, D.M., DeMayo, F.J., Katzenellenbogen, B.S., Lydon, J.P. and O’Malley, B.W. (2008) A repressive role for prohibitin in estrogen signaling. Molecular Endocrinology, 22, 344-360. http://dx.doi.org/10.1210/me.2007-0400