Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues

Ken-Ichirou Morohashi, Hiroshi Iida, Masatoshi Nomura, Osamu Hatano, Shin Ichiro Honda, Toshio Tsukiyama, Ohtsura Niwa, Takayuki Hara, Akira Takakusu, Yosaburo Shibata, Tsuneo Omura

Research output: Contribution to journalArticle

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Abstract

Ad4BP, a zinc finger DNA-binding protein, was identified as a transcription factor regulating steroidogenic P-450 genes in a cAMP-dependent manner. Immunochemical and immunohistochemical studies with steroidogenic tissues, adrenal, ovary, and testis, were performed using the antiserum to Ad4BP. Ad4BP was expressed to the same extent in the three zones of the adrenal cortex. Immunohistochemical examination of ovarian follicle and corpus luteum showed the expression of Ad4BP. The granulosa and thecal cells, the two distinct types of the steroidogenic cells in the follicle, gave Ad4BP signals, which were stronger than in the latter cells than in the former. Immunoblot analyses of mature and regressed corpora lutea indicated a parallel expression of Ad4BP and side-chain cleavage P-450, and both proteins significantly decreased in the regressed tissues. Leydig cells surrounding seminiferous tubules gave clear immunostaining signals for Ad4BP. ELP, a mammalian counterpart of Drosophila FTZ-F1 detected in EC cells, and are isoforms transcribed from the same gene. The Ad4BP and ELP forms recognize same nucleotide sequences. Reverse transcriptase-polymerase chain reaction with specific primers for ELP revealed that steroidogenic tissues contained ELP as well as Ad4BP. The effects of the two proteins on the transcription of the CYP11B gene were compared using the expression vectors of Ad4BP and ELP. ELP did not activate transcription and showed a weak inhibitory effect on the Ad4BP-dependent transactivation of the CYP11B gene promoter when transfected simultaneously. A gel shift analysis using in vitro synthesized Ad4BP and ELP revealed that the binding activity of ELP is significantly weaker than that of Ad4BP.

Original languageEnglish
Pages (from-to)643-653
Number of pages11
JournalMolecular Endocrinology
Volume8
Issue number5
DOIs
Publication statusPublished - May 1 1994

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Corpus Luteum
Genes
Seminiferous Tubules
Ovarian Follicle
Leydig Cells
Granulosa Cells
Adrenal Cortex
Zinc Fingers
DNA-Binding Proteins
Electrophoretic Mobility Shift Assay
Reverse Transcriptase Polymerase Chain Reaction
Transcriptional Activation
Drosophila
Testis
Immune Sera
Ovary
Protein Isoforms
Proteins
Transcription Factors
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues. / Morohashi, Ken-Ichirou; Iida, Hiroshi; Nomura, Masatoshi; Hatano, Osamu; Honda, Shin Ichiro; Tsukiyama, Toshio; Niwa, Ohtsura; Hara, Takayuki; Takakusu, Akira; Shibata, Yosaburo; Omura, Tsuneo.

In: Molecular Endocrinology, Vol. 8, No. 5, 01.05.1994, p. 643-653.

Research output: Contribution to journalArticle

Morohashi, K-I, Iida, H, Nomura, M, Hatano, O, Honda, SI, Tsukiyama, T, Niwa, O, Hara, T, Takakusu, A, Shibata, Y & Omura, T 1994, 'Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues', Molecular Endocrinology, vol. 8, no. 5, pp. 643-653. https://doi.org/10.1210/me.8.5.643
Morohashi, Ken-Ichirou ; Iida, Hiroshi ; Nomura, Masatoshi ; Hatano, Osamu ; Honda, Shin Ichiro ; Tsukiyama, Toshio ; Niwa, Ohtsura ; Hara, Takayuki ; Takakusu, Akira ; Shibata, Yosaburo ; Omura, Tsuneo. / Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues. In: Molecular Endocrinology. 1994 ; Vol. 8, No. 5. pp. 643-653.
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