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Horm Metab Res 2003; 35(5): 273-278
DOI: 10.1055/s-2003-41301
Original Basic
© Georg Thieme Verlag Stuttgart · New York

Pituitary Homeobox Factor 1, A Novel Transcription Factor in the Adrenal Regulating Steroid 11ß-hydroxylase

N.  Hiroi 1, 4 , T.  Kino 1 , M.  Bassett 5 , W.  E.  Rainey 5 , M.  Phung 1 , M.  Abu-Asab 2 , T.  Fojo 3 , P.  Briata 6, 7 , G.  P.  Chrousos 1 , S.  R.  Bornstein 1, 4
  • 1 Pediatric and Reproductive Endocrinology Branch, National Institute of Child Health and Human Development
  • 2 Laboratory of Pathology
  • 3 Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
  • 4 Department of Endocrinology, Heinrich Heine University of Düsseldorf, Germany
  • 5 Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, Texas, USA
  • 6 School and Department of Medicine, Howard Hughes Medical Institute, University of California, San Diego, La Jolla California, USA
  • 7 Istituto Nazionale Per La Ricerca Suc Cancro, Genova, Italy
This work was presented in part at the 84th annual meeting of the Endocrine Society.
Further Information

Publication History

Received 5 November 2002

Accepted after Revision 21 January 2003

Publication Date:
13 August 2003 (online)

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Abstract

Pituitary homeobox 1 (Ptx1/Pitx1) is a homeodomain-containing transcription factor present throughout pituitary development. Ptx1/Pitx1 interacts with steroidogenic factor 1 (SF-1) in the regulation of pituitary gene expression. SF-1 also plays a critical role in the transcription of enzymes involved in adrenal steroidogenesis. Therefore, we analyzed the presence and role of Ptx1/Pitx1 in human adrenal cortex. Both Ptx1/Pitx1 and SF-1 mRNA were expressed in the human adrenal gland, and immuno-electron microscopy demonstrated the presence of Ptx1/Pitx1 protein in the nucleus of adrenocortical cells. Computer analysis revealed the presence of Ptx1/Pitx1 signal sequences within the promoter region of human 11β hydroxylase (hCYP11B1). To examine the role of Ptx1/Pitx1 in the regulation of the genes, we prepared reporter constructs using the 5’-flanking DNA of the hCYP11B1 gene and transfected them into Y-1 mouse adrenocortical cells, HeLa and CV-1 cells. Ptx1/Pitx1 stimulation of hCYP11B1 reporter activity (3-fold over basal) in Y-1 cells was equal to that observed with SF-1. The hCYP11B1 promoter activity in Y-1 cells was not synergistically increased by co-transfection with both Ptx1/Pitx1 and SF-1. Both basal and ACTH-stimulated hCYP11B1 reporter activities in Y-1 cells were increased by co-transfection with either Ptx1/Pitx1 or SF-1 expression vectors. In contrast, co-transfection with both Ptx1/Pitx1 and SF-1 synergistically increased hCYP11B1 promoter activity in HeLa and CV-1 cells (5-fold and 20-fold over basal, respectively). In conclusion, this study represents the first demonstration for a role of Ptx1/Pitx1 in the regulation of transcription of enzymes involved in adrenal steroidogenesis.

Key words

Pituitary Homeobox Factor 1 Ptx1/Pitx1 - 11β-hydroxylase - Steroidogenic Factor 1 (SF-1)

Reference

  • 1 Lamonerie T, Tremblay J J, Lanctot C, Therrien M, Gauthier Y, Drouin J. Ptx1, a bicoid-related homeo box transcription factor involved in transcription of the pro-opiomelanocortin gene.  Genes Dev. 1996;  10 1284-1295
    PubMedSearch in Google Scholar
  • 2 Lanctot C, Gauthier Y, Drouin J. Pituitary homeobox 1 (Ptx1) is differentially expressed during pituitary development.  Endocrinology. 1999;  140 1416-1422
    CrossrefPubMedSearch in Google Scholar
  • 3 Tremblay J J, Lanctot C, Drouin J. The pan-pituitary activator of transcription, Ptx1 (pituitary homeobox 1), acts in synergy with SF-1 and Pit1 and is an upstream regulator of the Lim-homeodomain gene Lim3/Lhx3.  Mol Endocrinol. 1998;  12 428-441
    CrossrefPubMedSearch in Google Scholar
  • 4 Poulin G, Turgeon B, Drouin J. NeuroD1/beta2 contributes to cell-specific transcription of the proopiomelanocortin gene.  Mol Cell Biol. 1997;  17 6673-6682
    PubMedSearch in Google Scholar
  • 5 Tremblay J J, Marcil A, Gauthier Y, Drouin J. Ptx1 regulates SF-1 activity by an interaction that mimics the role of the ligand-binding domain.  EMBO J. 1999;  18 3431-3441
    CrossrefPubMedSearch in Google Scholar
  • 6 Lala D S, Rice D A, Parker K L. Steroidogenic factor I, a key regulator of steroidogenic enzyme expression, is the mouse homolog of fushi tarazu-factor I.  Mol Endocrinol. 1992;  6 1249-1258
    CrossrefPubMedSearch in Google Scholar
  • 7 Morohashi K, Zanger U M, Honda S, Hara M, Waterman M R, Omura T. Activation of CYP11A and CYP11B gene promoters by the steroidogenic cell-specific transcription factor, Ad4BP.  Mol Endocrinol. 1993;  7 1196-1204
    CrossrefPubMedSearch in Google Scholar
  • 8 Ikeda Y, Shen W H, Ingraham H A, Parker K L. Developmental expression of mouse steroidogenic factor-1, an essential regulator of the steroid hydroxylases.  Mol Endocrinol. 1994;  8 654-662
    CrossrefPubMedSearch in Google Scholar
  • 9 Achermann J C, Ito M, Hindmarsh P C, Jameson J L. A mutation in the gene encoding steroidgenic factor-1 cause reversal and adrenal failure in humans.  Nat Genet. 1999;  22 125-126
    CrossrefPubMedSearch in Google Scholar
  • 10 Sugawara T, Kiriakidou M, McAllister J M, Kallen C B, Strauss J F. 3rd Multiple steroidogenic factor 1 binding elements in the human steroidogenic acute regulatory protein gene 5’-flanking region are required for maximal promoter activity and cyclic AMP responsiveness.  Biochemistry. 1997;  36 7249-7255
    CrossrefPubMedSearch in Google Scholar
  • 11 Leers-Sucheta S, Morohashi K, Mason J I, Melner M H. Synergistic activation of the human type II 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase promoter by the transcription factor steroidogenic factor-1/adrenal 4-binding protein and phorbol ester.  J Biol Chem. 1997;  272 7960-7967
    CrossrefPubMedSearch in Google Scholar
  • 12 Rainey W E. Adrenal zonation: clues from 11beta-hydroxylase and aldosterone synthase.  Mol Cell Endocrinol. 1999;  151 151-160
    CrossrefPubMedSearch in Google Scholar
  • 13 Hiroi N, Chrousos G P, Kohn B, Bonat S, White A, Bornstein S R. Adrenocortical-pituitary hybrid tumor causing Cushing’s syndrome.  J Clin Endocrinol Metab. 2001;  86 2631-2637
    CrossrefPubMedSearch in Google Scholar
  • 14 Willenberg H S, Stratakis C A, Marx C, Ehrhart-Bornstein M, Chrousos G P, Bornstein S R. Aberrant interleukin-1 receptors in a cortisol-secreting adrenal adenoma causing Cushing’s syndrome.  N Engl J Med. 1998;  339 27-31
    CrossrefPubMedSearch in Google Scholar
  • 15 Schreiber E, Matthias P, Muller M M, Schaffner W. Rapid detection of octamer binding proteins with ‘mini-extract,’ prepared from a small number of cells.  Nucleic Acids Res. 1989;  17 6419
    PubMedSearch in Google Scholar
  • 16 Wang X L, Bassett M, Zhang Y, Yin S, Clyne C, White P C, Rainey W E. Transcriptional regulation of human 11beta-hydroxylase (hCYP11B1).  Endocrinology. 2000;  141 3587-3594
    CrossrefPubMedSearch in Google Scholar
  • 17 Rainey W E, Mrotek J J. Adrenal cortex tumors. In: Masters JRW, Passon B (eds) Human Cell Culture: Human Cancer Cell Lines. Kluwer Academic Press (Great Britain) 1998: 123-135
    Search in Google Scholar
  • 18 Szeto D P, Ryan A K, O’Connell S M, Rosenfeld M G. P-OTX: a PIT-1-interacting homeodomain factor expressed during anterior pituitary gland development.  Proc Natl Acad Sci USA. 1996;  93 7706-7710
    CrossrefPubMedSearch in Google Scholar
  • 19 Ikeda Y, Swain A, Weber T J, Hentges K E, Zanaria E, Lalli E, Tamai K T, Sassone-Corsi P, Lovell-Badge R, Camerino G, Parker K L. Steroidogenic factor 1 and Dax-1 colocalize in multiple cell lineages: potential links in endocrine development.  Mol Endocrinol. 1996;  10 1261-1272
    CrossrefPubMedSearch in Google Scholar
  • 20 Fernandez P M, Brunel F, Jimenez M A, Saez J M, Cereghini S, Zakin M M. Nuclear receptors Nor1 and NGFI-B/Nur77 play similar, albeit distinct, roles in the hypothalamo-pituitary-adrenal axis.  Endocrinology. 2000;  141 2392-2400
    CrossrefPubMedSearch in Google Scholar
  • 21 Lanctot C, Lamolet B, Drouin J. The bicoid-related homeoprotein Ptx1 defines the most anterior domain of the embryo and differentiates posterior from anterior lateral mesoderm.  Development. 1997;  124 2807-2817
    PubMedSearch in Google Scholar
  • 22 Tremblay J J, Goodyer C G, Drouin J. Transcriptional properties of Ptx1 and Ptx2 isoforms.  Neuroendocrinology. 2000;  71 277-286
    CrossrefPubMedSearch in Google Scholar
  • 23 Gage P J, Suh H, Camper S A. The bicoid-related Pitx gene family in development.  Mamm Genome. 1999;  10 197-200
    CrossrefPubMedSearch in Google Scholar
  • 24 Szeto D P, Rodriguez-Esteban C, Ryan A K, O’Connell S M, Liu F, Kioussi C, Gleiberman A S, Izpisua-Belmonte J C, Rosenfeld M G. Role of the Bicoid-related homeodomain factor Pitx1 in specifying hindlimb morphogenesis and pituitary development.  Genes Dev. 1999;  13 484-494
    PubMedSearch in Google Scholar
  • 25 Ryan A K, Rosenfeld M G. POU domain family values: flexibility, partnerships, and develpomental codes.  Gene Dev. 1997;  11 1207-1225
    PubMedSearch in Google Scholar
  • 26 Rosenfeld M G, Bach I, Erkman L, Li P, Lin C, Lin S, McEvilly R, Ryan A K, Rhodes S, Schnnemann M, Scully K. Transcriptional control of cell phenotypes in thye neuroendocrine system.  Recent Prog Horm Res. 1996;  51 217-238
    PubMedSearch in Google Scholar
  • 27 Andersen B, Rosenfeld M G. Pit-1 determines cell types during development of the anterior pituitary gland. A model for transcriptional regulation of cell phenotypes In: mammalian organogenesis.  J Biol Chem. 1994;  269 29 335-29 338
    PubMedSearch in Google Scholar
  • 28 Nachtigal M W, Hirokawa Y, Enyeart-VanHouten D L, Flanagan J N, Hammer G D, Ingraham H A. Wilms’ tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex-specific gene expression.  Cell. 1998;  93 445-454
    CrossrefPubMedSearch in Google Scholar

N. Hiroi, M. D.

First Department of Internal Medicine · Toho University School of Medicine ·

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