Abstract
Carbonic anhydrase III (CAIII) is an abundant muscle protein characteristic of adult type-1, slow-twitch, muscle fibers. We demonstrate that CAIII is not confined to mature muscle but is also expressed in cultured myogenic cells that were originally derived from adult and fetal limb muscle (G8 and C2C12) and by azacytidine treatment of 10T1/2 fibroblasts (23A2). Transcripts may accumulate in these cells to levels that correspond to 6.5% of that found in mature muscle. CAIII is expressed in mononucleate myoblasts and is abundant in those that preferentially fuse to form myotubes, and these findings contrast with those for many other muscle genes whose transcripts only accumulate on or after terminal differentiation. Preliminary promoter-function assays by transfection shows that 2.8 kb of sequence flanking the 5′ end of the human CAIII gene efficiently promotes transcription of the bacterial chloramphenicol acetyltransferase gene in myogenic cells. However, none of the sequences within this region are sufficient to confer muscle-specific expression. Removal of sequences 5′ to −715 bp leads to a major loss of transcriptional activity of the CAIII promoter. These results imply that the proximal CAIII promoter, which includes a putative CArG box and four potential MyoD binding sites, is not adequate for either myoblast-specific or maximal transcription.
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Literature cited
Jeffery, S., Edwards, Y., and Carter, N. (1980).Biochem. Genet. 18843–849.
Shiels, A., Jeffery, S., Wilson, C., and Carter, N. (1984).Biochem. J. 218281–284.
Jeffery, S., Carter, N., and Smith, A. (1986).J. Histochem. Cytochem. 34513–516.
Brownson, C., Isenberg, H., Brown, W., Salmons, S., and Edwards, Y. (1988).Muscle Nerve 111183–1189.
Gros,G., Ganhoff,F., Scheid, P., Siffert, W., Teske, W., and Kruger, D. (1984).Pflügers Arch. 400:R58.
Fremont, P., Lazure, C., Tremblay, R.R., Chretien, M., and Rogers, P.A. (1987).Biochem. Cell Biol. 65790–797.
Tashian, R.E. (1989).Bioessays 10186–192.
Jeffery, S., Wilson, C., Mode, A., Gustafsson, J., and Carter, N. (1986).J. Endocrinol. 110123–126.
Lloyd, J., McMillan, S., Hopkinson, D., and Edwards, Y.H. (1986).Gene 41233–239.
Tweedie, S., and Edwards, Y. (1989).Biochem. Genet. 2717–30.
Hastings, K., and Emerson, C. (1982).Proc. Natl. Acad. Sci. U.S.A. 791553–1557.
Rosenberg, U.B., Kunz, G. Frischauf, A. Lehrach, H. Mahr R., Eppenberger, H.M., and Perriard J. (1982)Proc. Natl. Acad. Sci. USA 796589–6592.
Klamut, H.J., Gangopadhyay, S.B., Worton, R.G., and Ray, P.N. (1990).Mol. Pharmacol. 10193–205.
Weller, P.A., Price, M., Isenberg, H., Edwards, Y.H., and Jeffreys, A.J. (1986).Mol. Cell. Biol. 64539–4547.
Lloyd, J., Brownson, C., Tweedie, S., Charlton, J., and Edwards, Y.H. (1987).Genes Dev. 1594–602.
Edwards, Y.H. (1990). InThe Carbonic Anhydrases: Cellular Physiology & Molecular Genetics (ed.) Dodgson, S. (Plenum Press, New York).
Edwards, Y.H., Charlton, J., and Brownson, C. (1988).Gene 71473–481.
Colantuoni, V., Pirozzi, A., Blance, C., and Cortese, R. (1987).EMBO J. 6631–636.
Christian, C.N., and Nelson, P.G. (1977).Science 196995–998.
Yaffe, D., and Saxel, O. (1977).Nature 270725–727.
Blau, H.M., Chiu, C.-P., and Webster, C. (1983).Cell 321171–1180.
Konieczny, S.F., and Emerson, C.P., Jr., (1984).Cell 38791–800.
Reznikoff, C.A., Brankow, D.W., and Heidelberger, C. (1973).Cancer Res. 333231–3238.
Gillespie, G., Lloyd, J., Hopkinson, D.A., and Edwards, Y. (1984).J. Muscle Res. Cell Motil. 5457–464.
Edwards, Y., Lloyd, J., McMillan, S., and Benham, F. (1985).Mol. Cell. Biol. 52147–2149.
Frebourg, T., and Brison, O. (1988).Gene 65315–318.
Gorman, C.M., Merilino, G.T., Willingham, M.C., Pastan, I., and Howard, B.H. (1982).Proc. Natl. Acad. Sci. U.S.A. 796777–6781.
Gunning, P.L., Leavitt, J., Muscat, G., Ng, S.-Y., and Kedes, L. (1987).Proc. Natl. Acad. Sci. U.S.A. 844831–4835.
Gorman, C. (1985). DNA Cloning Vol 11 IRL Press Oxford, Washington DC pp. 143–190.
Richler, C., and Yaffe, D. (1970).Dev. Biol. 231–22.
Miller, J.B., and Stockdale, F.E. (1986).Proc. Natl. Acad. Sci. U.S.A. 833860–3864.
Sassoon, D., Lyons, G., Wright, W., Lin, V., Lassar, A., Weintraub, H., and Buckingham, M. (1989).Nature 341303–307.
Braun, T., Bober, E., Buschhausen-Denker, G., Kotz, S., Grzeschik, K.H., and Arnold, H.H. (1989).EMBO J. 83617–3625.
Pinney, D., Pearson-White, S., Konieczny, S., Latham, K., and Emerson, C. (1988).Cell 53781–793.
Miwa, T., and Kedes, L. (1987).Mol. Cell Biol. 72803–2813.
Lassar, A.B., Buskin, J.N., Lockshon, D., Davis, R.L., Apone, S., Hauschka, S.D., and Weintraub, H. (1989).Cell 58823–831.
Buskin, J.N., and Hauschka, S.D. (1989).Mol. Cell. Biol. 92627–2640.
Zenke, M., Grundström, T., Mattes, H., Wintezerith, M., Schatz, C., Wildeman, A., and Chambon, P. (1986).EMBO. J. 5387–397.
Jaynes, J., Johnson, J., Buskin, J., Gartside, C., and Hauschka, S. (1988).Mol. Cell Biol. 862–70.
Sternberg, E.A., Spizz, G., Perry, M., Vizard, D., Weil, T., and Olson, E.N. (1988).Mol. Cell Biol. 82896–2909.
Shapiro, L.H., Venta, P.J., and Tashian, R.E. (1987).Mol. Cell Biol. 74589–4593.
Bird, A.P. (1986).Nature 321209–213.
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Tweedie, S., Morrison, K., Charlton, J. et al. CAIII a marker for early myogenesis: Analysis of expression in cultured myogenic cells. Somat Cell Mol Genet 17, 215–228 (1991). https://doi.org/10.1007/BF01232818
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DOI: https://doi.org/10.1007/BF01232818