Cyclic AMP-responsive region of the human proopiomelanocortin (POMC) gene

Usui, Takeshi; Nakai, Yoshikatsu; Tsukada, Toshihiko; Fukata, Junichi; Nakaishi, Shigeo; Naitoh, Yoshiyuki; Imura, Hiroo

doi:10.1016/0303-7207(89)90123-8

Cited by 16 publications

(6 citation statements)

References 20 publications

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“…1983) but its promoter has been poorly characterized. Trans¬ fection studies with the human POMC promoter have long been hampered by its weak activity, requiring the establishment of stable cell lines (Usui et al 1989) or the use of a viral enhancer to force POMC transcription (Mishina et al 1982, Israel 8c Cohen 1985. In addition, the human POMC promoter has not been studied in a human cell line with endogenous POMC expression because no human equivalent of AtT-20 has been available.…”

Section: Resultsmentioning

confidence: 99%

Functional analysis of the human pro-opiomelanocortin promoter in the small cell lung carcinoma cell line DMS-79

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Leblond-Francillard²,

Raffin-Sanson³

et al. 1995

Journal of Molecular Endocrinology

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DMS-79 is a human cell line derived from a small cell lung carcinoma (SCLC), which expresses the pro-opiomelanocortin (POMC) gene. We took it as a model in which to study the mechanism of POMC gene expression in these tumors: precursor processing is altered and gene expression is essentially unresponsive to glucocorticoids. POMC gene structure appeared normal by Southern blot analysis, indicating that gene rearrangement was not responsible for its expression in DMS-79. Indeed, using transient expression of human POMC-luciferase fusion genes in DMS-79, we showed that (1) the normal human POMC promoter was functional in DMS-79, and (2) the same proximal promoter region (-417; + 21) produced the full transcriptional activity in DMS-79 and in the mouse pituitary cell line AtT-20. Progressive 5' deletion analysis revealed differences between AtT-20 and DMS-79: region (-611; -376) was active in AtT-20 and not in DMS-79, whereas region (-95; -161) was active in both cell lines and (-376; -417) was only active in DMS-79. The latter partially overlaps a motif homologous to the DE-2 rat element which confers the tissue-specific expression of POMC in AtT-20 cells; however, this motif had no effect in DMS-79. These data suggest that POMC gene transcription is achieved through a different set of transacting factors in DMS-79 and AtT-20. Altogether, our results provide evidence that DMS-79 is a valid model of tumors responsible for the ectopic ACTH syndrome and that the mechanism of POMC gene expression in these SCLC cells is different from that in pituitary cells.

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Section: Resultsmentioning

confidence: 99%

Functional analysis of the human pro-opiomelanocortin promoter in the small cell lung carcinoma cell line DMS-79

Picon

Leblond-Francillard²,

Raffin-Sanson³

et al. 1995

Journal of Molecular Endocrinology

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“…A palindromic consensus sequence for the cAMP response element (CRE; TGACGTCA) has been previously identified in many genes whose expression is modulated by cAMP (4)(5)(6)(7)(8)(9)(10). The corresponding binding protein (CREB) has been purified (11) and the cDNA encoding the CREB has been cloned (12,13).…”

Section: Embl Accession No X51428mentioning

confidence: 99%

Macromolecular interaction on a cAMP responsive region in the urokinase-type plasminogen activator gene: a role of protein phosphorylation

1990

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We have studied the regulation of urokinase-type plasminogen activator gene expression by cAMP in LLC-PK1 cells. We found a cAMP responsive region 3.4 kb upstream of the transcription initiation site, which comprised three protein-binding domains designated A, B, and C. Domains A and B both contain a sequence, TGACG, homologous to a consensus cAMP response element (CRE; TGACGTCA). Effective cAMP-mediated induction was achieved when these two domains were linked with domain C, which by itself did not confer cAMP responsiveness to a heterologous promoter nor contained CRE-like sequence, suggesting a functional cooperation among these domains. Results of competition studies using gel retardation and DNase I footprinting assays suggest that there is a protein-protein interaction between a CRE binding protein and a domain C binding protein. In gel retardation assays, binding of a nuclear protein to domains A and B was strongly augmented by addition of the catalytic subunit of cAMP-dependent protein kinase, whereas the protein binding to domain C was slightly inhibited, suggesting that protein phosphorylation is involved in the regulation of protein-DNA interaction.

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“…The plasmid pPOMCGALl was constructed by ligating the 3-kb Eco Rl-Hind III fragment from pPOMC-CATl (Usui et al, 1989), which contains 2.9 kb of the 5'-flanking region as well as 67 bp of exon 1 of the human POMC gene (Takahashi et al, 1981), and the 4.4-kb Hind lll-Pst I fragment from pCHHO, which contains the E. coli lacZ gene (Hall et al, 1983) as well as SV40 polyadenylation signals, into the vector pUC12 digested with Eco RI and Pst I. The plasmid pPOMCGAL2 was constructed by ligating the Pvu ll-Hind III fragment (approximately 770 bp) and the Hind Ill-Bam HI fragment (3.9 kb) from pPOMCGALl into the pBluescript SK+ (Stratagene) digested with Eco RV and Bam HI.…”

Section: Construction Offusion Genesmentioning

confidence: 99%

Functional Analysis of the Cell-Specific Enhancer in the Human Proopiomelanocortin Gene by β-Galactosidase Histochemical Staining

Tsukada

Nakai²,

Fukushima³

et al. 1994

DNA and Cell Biology

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Nucleotide sequences responsible for the cell-specific expression of the human proopiomelanocortin (POMC) gene were analyzed by histochemical staining of beta-galactosidase in culture cells transfected with chimeric genes containing the 5'-flanking regions of the human POMC gene fused to the Escherichia coli lacZ gene. The chimeric genes were stably introduced into various culture cells, including AtT-20 cells, which express the endogenous mouse POMC gene. Whereas the control gene containing the cytomegalovirus enhancer was expressed in all cell lines tested, only AtT-20 cells supported the efficient transcription of the gene containing 2.9 kb of the human POMC 5'-flanking region. These results indicate that the stable transfection-expression system utilizing the histochemical detection of the gene expression is a useful method for the analysis of cell-specific gene expression. These results have also confirmed that the trans-acting factors in mouse AtT-20 cells interact with the human POMC gene promoter region and activate the transcription of the gene. Deletion analysis has demonstrated that the profiles of the transcriptional activity of the various human POMC-lacZ fusion genes are similar to those of the rat POMC gene described previously. Comparison of the human and the rat 5'-flanking sequences revealed close homology in several regions, which might be involved in the efficient transcription of the POMC gene in AtT-20 cells.

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Cyclic AMP-responsive region of the human proopiomelanocortin (POMC) gene

Cited by 16 publications

References 20 publications

Functional analysis of the human pro-opiomelanocortin promoter in the small cell lung carcinoma cell line DMS-79

Functional analysis of the human pro-opiomelanocortin promoter in the small cell lung carcinoma cell line DMS-79

Macromolecular interaction on a cAMP responsive region in the urokinase-type plasminogen activator gene: a role of protein phosphorylation

Functional Analysis of the Cell-Specific Enhancer in the Human Proopiomelanocortin Gene by β-Galactosidase Histochemical Staining

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