Interaction between the Wilms tumour factor-1 element in the promoter of Amh and a downstream enhancer is required for a strong expression of the gene in pre-pubertal sertoli cells

David W. Dresser

The Ashworth Laboratory, The University of Edinburgh, Edinburgh, UK.

DOI: 10.4236/ajmb.2013.33022                

Abstract

Amh (anti-Müllerian hormone) is a single copy gene which is expressed strongly in Sertoli cells in the foetal testis and participates in the onset of sexual differentiation. Its promoter driving the expression of a reporter gene (d2EGFP) has been used to analyse the role of certain defined putative elements and a downstream enhancer element in gene expression. These experiments were carried out in vitro using a line of pre-pubertal mouse Sertoli cells, transienly transfected with circular DNA constructs with variously mutated promoter elements. A downstream enhancer element, situated immediately 3’ of the polyadenylation (PA) signal for Amh, has been inserted in an equivalent position in the d2EGFP construct. When the Amh promoter is unmodified, the downstream enhancer (DE) is positively associated with a large increase in EGFP expression. This is at least partly the consequence of an increased rate of expression by individual cells. Experiments using variously truncated Amh promoters indicate that an upstream region (-214 to -336) may play a minor role in facilitating enhancement. However mutation of the Wilms tumour factor-1 element, situated between the tata box and the start of translation, results in an almost complete suppression of enhancement.

Keywords

Mouse Cell Lines; In Vitro; Amh Promoter; SMAT; Pre-Pubertal Sertoli; Downstream Enhancer; Wilms Tumour Factor Element

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Josso, N., Picard, J.Y., Rey, R. and di Clemente, N. (2006) Testicular anti-Mullerian hormone: History, genetics, regulation and clinical applications. Pediatric Endocrinology Reviews, 3, 347-358.
[2]	Durlinger, A.L., Visser, J.A. and Themmen, A.P. (2002) Regulation of ovarian function: The role of anti-Mullerian hormone. Reproduction, 124, 601-609. doi:10.1530/rep.0.1240601
[3]	Koopman, P., Gubbay, J., Vivian, N., Goodfellow, P. and Lovell-Badge, R. (1991) Male development of chromosomally female mice transgenic for Sry. Nature, 351, 117-121. doi:10.1038/351117a0
[4]	Dresser, D.W. (2012) Mutated elements of a complex promoter (Amh) can help to demonstrate the role of certain elements in controlling differential gene expression. American Journal of Molecular Biology, 2, 351-358. doi:10.4236/ajmb.2012.24036
[5]	de Santa Barbara, P., Moniot, B., Poulat, F. and Berta, P. (2000) Expression and subcellular localization of SF-1, SOX9, WT1, and AMH proteins during early human testicular development. Developmental Dynamics, 217, 293-298. doi:10.1002/(SICI)1097-0177(200003)217:3<293::AID-DVDY7>3.0.CO;2-P
[6]	Oreal, E., Mazaud, S., Picard, J.Y., Magre, S. and Carre-Eusebe, D. (2002) Different patterns of anti-Mullerian hormone expression, as related to DMRT1, SF-1, WT1, GATA-4, Wnt-4, and Lhx9 expression, in the chick differentiating gonads. Developmental Dynamics, 225, 221-232. doi:10.1002/dvdy.10153
[7]	Arango, N.A., Lovell-Badge, R. and Behringer, R.R. (1999) Targeted mutagenesis of the endogenous mouse Mis gene promoter: In vivo definition of genetic pathways of vertebrate sexual development. Cell, 99, pp. 409-419. doi:10.1016/S0092-8674(00)81527-5
[8]	Münsterberg, A. and Lovell-Badge, R. (1991) Expression of the mouse anti-mullerian hormone gene suggests a role in both male and female sexual differentiation. Development, 113, 613-624.
[9]	Schepers, G., Wilson, M., Wilhelm, D. and Koopman, P. (2003) SOX8 is expressed during testis differentiation in mice and synergizes with SF1 to activate the Amh promoter in vitro. Journal of Biological Chemistry, 278, 28101-28108. doi:10.1074/jbc.M304067200
[10]	Dresser, D.W. and Guerrier, D. (2005) Candidate sertoli cell specific promoter element for a TGFbeta family member (Amh) and a 3’ UTR enhancer/repressor for the same gene. Gene, 363, 159-165. doi:10.1016/j.gene.2005.08.004
[11]	Anderson, A.A., Treves, S., Biral, D., Betto, R., Sandonà, D., Ronjat, M. and Zorzato, F. (2003) The novel skeletal muscle sarcoplasmic reticulum JP-45 protein. Molecular cloning, tissue distribution, developmental expression, and interaction with alpha 1.1 subunit of the voltagegated calcium channel. Journal of Biological Chemistry, 278, 39987-39989. doi:10.1074/jbc.M305016200
[12]	Belville, C., Jamin, S.P., Picard, J.Y., Josso, N. and di Clemente, N. (2005) Role of type I receptors for anti-Mullerian hormone in the SMAT-1 Sertoli cell line. Oncogene, 24, 4984-4992.
[13]	Dresser, D.W., Hacker, A., Lovell-Badge, R. and Guerrier, D. (1995) The genes for a spliceosome protein (Sap 62) and the anti-Mullerian Hormone are contiguous. Human Molecular Genetics, 4, 613-618. doi:10.1093/hmg/4.9.1613
[14]	Anderson, A.M., Weasner, B.M., Weasner, B.P. and Kumar, B.P. (2012) Dual transcriptional activities of SIX define their roles in normal and ectopic eye development. Development, 139, 991-1000. doi:10.1242/dev.077255
[15]	Miyamota, Y., Taniguchi, H., Hamel, F., Silversides, D.W. and Viger, R. (2008) A GATA4/WT1 cooperation regulates transcription of genes required for mammalian sex derermination and differentiation. BMC Molecular Biology, 9, 44-62. doi:10.1186/1471-2199-9-44
[16]	Viger, R.S., Taniguchi, H., Robert, N.M. and Tremblay, J.J. (2004) Role of the GATA family of transcription factors in andrology. Journal of Andrology, 25, 441-452.
[17]	Klattig, J., Sierig, R., Kruspe, D., Besenbeck, B. and Englert, C. (2007) Wilms’ tumor protein Wt1 is an activator of the anti-Mullerian hormone receptor gene Amhr2. Molecular and Cellular Biology, 27, 4355-4364. doi:10.1128/MCB.01780-06
[18]	Guittot, S.M., Tetu, A., Legault, E., Pilon, N., Silversides, D.W. and Viger, R.S. (2007) The proximal Gata4 promoter directs reporter gene expression to sertoli cells during mouse gonadal development. Biology of Reproduction, 76, 85-95. doi:10.1095/biolreprod.106.055137
[19]	Sekido, R. and Lovell-Badge, R. (2008) Sex determination involves synergistic action of SRY and SF1 on a specific Sox9 enhancer. Nature, 453, 930-934. doi:10.1038/nature06944
[20]	Merritt, C., Rasojoson, D., Ko, D. and Seydoux, G. (2008) 3’ UTR are the primary regulators of gene expression in the C. elegansgermline. Current Biology, 14, 1476-1482. doi:10.1016/j.cub.2008.08.013
[21]	Derrigo, M., Cestelli, A., Savettieri, G. and diLiegro, I. (2000) RNA-protein interactrions in the control of stability and localization of messenger RNA. International Journal of Molecular Medicine, 5, 111-123.
[22]	Gallie, D.R. (1991) The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes & Development, 5, 2108-2116. doi:10.1101/gad.5.11.2108
[23]	White, S. and Sinclair, A. (2012) The molecular basis of gonadal development and disorders of sex development. Chapter 1 in J. M. Hutson, et al., Eds., Disorders of Sex Development, Springer, Berlin, pp. 1-9. doi:10.1007/978-3-642-22964-0_1
[24]	Furuhata, A., Murakami, M., Ito, H., Yoshida, K., Sobue, S., Kikuchi, R., Iwasaki, T., Takagi, A., Kojima, T., Suzuki, M., Abe, A. and Murate, T. (2009) GATA-1 and GATA-2 binding to 3’ enhancer of WT1 gene is essential for its transcription in acuteleukemia and solid tumor cells. Leukemia, 23, 1270-1277. doi:10.1038/leu.2009.13