- Research Article
Characterization of the 5′-flanking region of the mouse asparagine-linked glycosylation 12 homolog gene
Cellular & Molecular Biology Letters volume 18, pages 315–327 (2013)
Recently, we characterized multiple roles of the endoplasmic reticulum stress responsive element (ERSE) in the promotion of a unique headto-head gene pair: mammalian asparagine-linked glycosylation 12 homolog (ALG12) and cysteine-rich with EGF-like domains 2 (CRELD2). This bidirectional promoter, which consists of fewer than 400 base pairs, separates the two genes. It has been demonstrated that the ALG12 promoter shows less transcriptional activity through ERSE, but its basic regulatory mechanism has not been characterized. In this study, we focused on well-conserved binding elements for the transcription factors for ATF6, NF-Y and YY1 and the Sp1 and Ets families in the 5’-flanking region of the mouse ALG12 gene. We characterized their dominant roles in regulating ALG12 promoter activities using several deletion and mutation luciferase reporter constructs. The ALG12 gene is expressed in three distinct cell lines: Neuro2a, C6 glioma and HeLa cells. The reporter activity in each cell line decreased similarly with serial deletions of the mouse ALG12 promoter. Mutations in the ERSE and adjacent NF-Y-binding element slightly affected reporter activity. Each of the mutations in the GC-rich sequence and YY1-binding element reduced ALG12 promoter activity, and the combination of these mutations additively decreased reporter activity. Each mutation in the tandem-arranged Ets-family consensus sequences partially attenuated ALG12 promoter activity, and mutations of all three Ets-binding elements decreased promoter activity by approximately 40%. Mutation of the three conserved regulatory elements (GC-rich, YY1 and Ets) in the ALG12 promoter decreased reporter activity by more than 90%. Our results suggest that the promoter activity of the mouse ALG12 gene is regulated in a similar manner in the three cell lines tested in this study. The well-conserved consensus sequences in the promoter of this gene synergistically contribute to maintaining basal gene expression.
asparagine-linked glycosylation 12 homolog
activating transcription factor 6
congenital disorders of glycosylation
cysteine-rich with EGF-like domains 2
ER stress response element
v-ets erythroblastosis virus E26 oncogene homolog
nuclear transcription factor Y
Yin Yang 1
Grubenmann, C.E., Frank, C.G., Kjaergaard, S., Berger, E.G., Aebi, M. and Hennet, T. ALG12 mannosyltransferase defect in congenital disorder of glycosylation type lg. Hum. Mol. Genet. 11 (2002) 2331–2339.
Chantret, I., Dupré, T., Delenda, C., Bucher, S., Dancourt, J., Barnier, A., Charollais, A., Heron, D., Bader-Meunier, B., Danos, O., Seta, N., Durand, G., Oriol, R., Codogno, P. and Moore, S.E. Congenital disorders of glycosylation type Ig is defined by a deficiency in dolichyl-Pmannose: Man7GlcNAc2-PP-dolichyl mannosyltransferase. J. Biol. Chem. 277 (2002) 25815–25822.
Burda, P., Jakob, C.A., Beinhauer, J., Hegemann, J.H. and Aebi, M. Ordered assembly of the asymmetrically branched lipid-linked oligosaccharide in the endoplasmic reticulum is ensured by the substrate specificity of the individual glycosyltransferases. Glycobiology 9 (1999) 617–625.
Hoseki, J., Ushioda, R. and Nagata, K. Mechanism and components of endoplasmic reticulum-associated degradation. J. Biochem. 147 (2010) 19–25.
Adachi, N. and Lieber, M.R. Bidirectional gene organization: a common architectural feature of the human genome. Cell 109 (2002) 807–809.
Trinklein, N.D., Aldred, S.F., Hartman, S.J., Schroeder, D.I., Otillar, R.P. and Myers, R.M. An abundance of bidirectional promoters in the human genome. Genome Res. 14 (2004) 62–66.
Li, Y.Y., Yu, H., Guo, Z.M., Guo, T.Q., Tu, K. and Li, Y.X. Systematic analysis of head-to-head gene organization: evolutionary conservation and potential biological relevance. PLoS Comput. Biol. 2 (2006) e74.
Oh-hashi, K., Koga, H., Ikeda, S., Shimada, K., Hirata, Y. and Kiuchi, K. Role of an ER stress response element in regulating the bidirectional promoter of the mouse CRELD2 — ALG12 gene pair. BMC Genomics 11 (2010) 664.
Oh-hashi, K., Koga, H., Ikeda, S., Shimada, K., Hirata, Y. and Kiuchi, K. CRELD2 is a novel endoplasmic reticulum stress-inducible gene. Biochem. Biophys. Res. Commun. 387 (2009) 504–510.
Oh-hashi, K., Kunieda, R., Hirata, Y. and Kiuchi, K. Biosynthesis and secretion of mouse cysteine-rich with EGF-like domains 2. FEBS Lett. 585 (2011) 2481–2487.
Ortiz, J.A., Castillo, M., del Toro, E.D., Mulet, J., Gerber, S., Valor, L.M., Sala, S., Sala, F., Gutiérrez, L.M. and Criado, M. The cysteine-rich with EGF-like domains 2 (CRELD2) protein interacts with the large cytoplasmic domain of human neuronal nicotinic acetylcholine receptor α4 and β2 subunits. J. Neurochem. 95 (2005) 1585–1596.
Zhu, C., Johansen, F.E. and Prywes, R. Interaction of ATF6 and serum response factor. Mol. Cell. Biol. 17 (1997) 4957–4966.
Haze, K., Yoshida, H., Yanagi, H., Yura, T. and Mori, K. Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Mol. Biol. Cell. 10 (1999) 3787–3799.
Mantovani, R. The molecular biology of the CCAAT-binding factor NF-Y. Gene 239 (1999) 15–27.
Dynan, W.S. and Tjian, R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell 35 (1983) 79–87.
Kaczynski, J., Cook, T. and Urrutia, R. Sp1- and Krüppel-like transcription factors. Genome Biol. 4 (2003) 206.
Gordon, S., Akopyan, G., Garban, H. and Bonavida, B. Transcription factor YY1: structure, function, and therapeutic implications in cancer biology. Oncogene 25 (2006) 1125–1142.
Deng, Z., Cao, P., Wan, M.M. and Sui, G. Yin Yang 1: a multifaceted protein beyond a transcription factor. Transcription 1 (2010) 81–84.
Wasylyk, B., Hahn, S.L. and Giovane, A. The Ets family of transcription factors. Eur. J. Biochem. 211 (1993) 7–18
Oettgen, P. Regulation of vascular inflammation and remodeling by ETS factors. Circ. Res. 99 (2006) 1159–1166.
Kadonaga, J.T., Carner, K.R., Masiarz, F.R. and Tjian, R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell 51 (1987) 1079–1090.
Marin, M., Karis, A., Visser, P., Grosveld, F. and Philipsen, S. Transcription factor Sp1 is essential for early embryonic development but dispensable for cell growth and differentiation. Cell 89 (1997) 619–628.
Donohoe, M.E., Zhang, X., McGinnis, L., Biggers, J., Li, E. and Shi, Y. Targeted disruption of mouse Yin Yang 1 transcription factor results in periimplantation lethality. Mol. Cell. Biol. 19 (1999) 7237–7244.
Yang, W.M., Inouye, C., Zeng, Y., Bearss, D. and Seto, E. Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3. Proc. Natl. Acad. Sci. USA 93 (1996) 12845–12850.
Sui, G., Affarel, B., Shi, Y., Brignone, C., Wall, N.R., Yin, P., Donohoe, M., Luke, M.P., Calvo, D., Grossman, S.R. and Shi, Y. Yin Yang 1 is a negative regulator of p53. Cell 117 (2004) 859–872.
Lie-Venema, H., Gittenberger-de Groot, A.C., van Empel, L.J., Boot, M.J., Kerkdijk, H., de Kant, E. and DeRuiter, M.C. Ets-1 and Ets-2 transcription factors are essential for normal coronary and myocardial development in chicken embryos. Circ. Res. 92 (2003) 749–756.
Seth, A. and Watson, D.K. ETS transcription factors and their emerging roles in human cancer. Eur. J. Cancer 41 (2005) 2462–2478.
Jinnin, M., Ihn, H., Asano, Y, Yamane, K, Trojanowska, M. and Tamaki, K. Tenascin-C upregulation by transforming growth factor-β in human dermal fibroblasts involves Smad3, Sp1, and Ets1. Oncogene 23 (2004) 1656–1667.
Lee, C.G., Kwon, H.K., Sahoo, A., Hwang, W., So, J.S., Hwang, J.S., Chae, C.S., Kim, G.C., Kim, J.E., So, H.S., Hwang, E.S., Grenningloh, R., Ho, I.C. and Im, S.H. Interaction of Ets-1 with HDAC1 represses IL-10 expression in Th1 cells. J. Immunol. 188 (2012) 2244–2253.
Enya, K., Hayashi, H., Takii, T., Ohoka, N., Kanata, S., Okamoto, T. and Onozaki, K. The interaction with Sp1 and reduction in the activity of histone deacetylase 1 are critical for the constitutive gene expression of IL-1α in human melanoma cells. J. Leukoc. Biol. 83 (2008) 190–199.
Haeuptle, M.A. and Hennet, T. Congenital disorders of glycosylation: an update on defects affecting the biosynthesis of dolichol-linked oligosaccharides. Hum. Mutat. 30 (2009) 1628–1641.
Tan, N.Y., Khachigian, L.M. Sp1 phosphorylation and its regulation of gene transcription. Mol. Cell. Biol. 29 (2009) 2483–2488.
Wasylyk, B., Hagman, J. and Gutierrez-Hartmann, A. Ets transcription factors: nuclear effectors of the Ras-MAP-kinase signaling pathway. Trends Biochem. Sci. 23 (1998) 213–216.
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Oh-Hashi, K., Tejima, T., Hirata, Y. et al. Characterization of the 5′-flanking region of the mouse asparagine-linked glycosylation 12 homolog gene. Cell Mol Biol Lett 18, 315–327 (2013). https://doi.org/10.2478/s11658-013-0091-2
- Ets family
- Sp1 family