Characterization of the rat Class 3 aldehyde dehydrogenase gene promoter

Xie, Y; Takimoto, Koichi; Pitot, Henry C.; Miskimins, W. Keith; Lindahl, Ronald

doi:10.1093/nar/24.21.4185

Cited by 20 publications

(13 citation statements)

References 32 publications

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“…DNase I footprinting experiments indicate that AP1 and Sp1 are involved in expression of the rat ALDH3 gene in hepatoma cells (35). AP2 and Sp1 are involved in the regulation of keratin gene expression in the cornea.…”

Section: Resultsmentioning

confidence: 96%

“…Although cultured hepatoma cells have been used to study induced expression of the rat ALDH3 gene (33)(34)(35), examination of available primary and simian virus 40-transformed corneal epithelial cells showed that the endogenous ALDH3 gene is not expressed in cultured corneal cells (data not shown). Similarly, ALDH3 activity declines in intact rat corneas placed in culture (25).…”

Section: Resultsmentioning

confidence: 99%

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Aldehyde dehydrogenase class 3 expression: Identification of a cornea-preferred gene promoter in transgenic mice

Kays

Piatigorsky

1997

Proc. Natl. Acad. Sci. U.S.A.

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Aldehyde dehydrogenase class 3 (ALDH3) constitutes 20-40% of the total water-soluble proteins in the mammalian cornea. Here, we show by Northern blot analysis that ALDH3 expression in the mouse is at least 500-fold higher in the cornea than in any other tissue examined, with very low levels of expression detected in the stomach, urinary bladder, ocular lens, and lung. Histochemical localization reveals that this exceptional level of expression in the mouse cornea occurs in the anterior epithelial cells and that little ALDH3 is present in the keratocytes or corneal endothelial cells. A 13-kbp mouse ALDH3 promoter fragment containing >12 kbp of the 5 flanking sequence, the 40-bp untranslated first exon, and 29 bp of intron 1 directed cat reporter gene expression to tissues that express the endogenous ALDH3 gene, except that transgene promoter activity was higher in the stomach and bladder than in the cornea. By contrast, when driven by a 4.4-kbp mouse ALDH3 promoter fragment [1,050-bp 5 flanking region, exon 1, intron 1 (3.4 kbp), and 7 bp of exon 2] expression of the cat reporter gene was confined to the corneal epithelial cells, except for very low levels in the liver, effectively reproducing the corneal expression pattern of the endogenous ALDH3 gene. These results indicate that tissue-specific expression of ALDH3 is determined by positive and negative elements in the 5 flanking region of the gene and suggests putative silencers located in intron 1. We demonstrate regulatory sequences capable of directing cornea-specific gene expression, affording the opportunity for genetic engineering in this transparent tissue.

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Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Aldehyde dehydrogenase class 3 expression: Identification of a cornea-preferred gene promoter in transgenic mice

Kays

Piatigorsky

1997

Proc. Natl. Acad. Sci. U.S.A.

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“…A series of deletion mutants of the proximal 5Ј-flanking region of aldh3 were prepared by polymerase chain reaction as described previously (6), namely pI-3, pI-4, pI-5, pI-6, and pI-8. Construct p⌬(Ϫ1054/Ϫ392)ALDH was produced by treating pALDH3.5 with the restriction enzyme PstI to remove a portion of the flanking region from Ϫ1057 bp to Ϫ392 bp relative to the transcription start site; the resulting plasmid was gel purified and religated to yield p⌬(Ϫ1057/Ϫ392)ALDH.…”

Section: Methodsmentioning

confidence: 99%

“…The data shown are representative of three separate experiments where the effect of the H8 was to cause a 2.7-3.0-fold increase in aldh3 mRNA levels; dibutyryl cAMP (DiBC) or forskolin (FOR) decreased both BA induction and the H8 potentiation of BA induction by 60 -75%, in all cases. (1,5,6,30) have developed a highly inducible 5Ј-flanking construct containing 3.5 kb of upstream sequence of the rat aldh3 gene in a chloramphenicol acetyltransferase (CAT) reporter gene. Basal level expression of this gene is cell type-specific, and liver displays very low basal expression (1, 2).…”

Section: Figmentioning

confidence: 99%

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cAMP-dependent Negative Regulation of Rat Aldehyde Dehydrogenase Class 3 Gene Expression

Xiao

Falkner

Xie

et al. 1997

Journal of Biological Chemistry

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We investigated the inhibitory effects of intracellular cyclic adenosine monophosphate (cAMP) levels in regulating class 3 aldehyde dehydrogenase (aldh3) gene expression using cultures of primary rat hepatocytes and transient transfection experiments with HepG2 cells. In addition to regulation by an Ah receptor-dependent mechanism, expression of many members of the Ah gene battery have been shown to be negatively regulated. As was seen for the cytochrome P450 (cyp1A1) gene, aldh3 is transcriptionally inducible by polycyclic aromatic hydrocarbons (PAH), and this induction involving function of the arylhydrocarbon (Ah) receptor is inhibited by the protein kinase C (PKC) inhibitors, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine di-HCl (H7) and staurosporine. However, PAH induction of ALDH-3 activity, protein, and mRNA was potentiated 2-4-fold by addition of the protein kinase A (PKA) inhibitors, N-(2-(methylamino)ethyl)-5-isoquinolinesulfonamide di-HCl (H8) and N-(2-guanidinoethyl)-5-isoquinolinesulfonamide HCl (HA1004). These PKA inhibitors had no effect on the PAH induction of the cyp1A1. Protein kinase A activity of cultured hepatocytes was specifically inhibited by H8 and HA1004 in a concentration-dependent manner, but not by H7, and there was an inverse correlation observed between potentiation of PAH-induced aldh3 gene expression and inhibition of specific PKA activity by the PKA inhibitors. The cAMP analog dibutyryl cAMP, the adenylate cyclase activator forskolin, and the protein phosphatase 1 and 2A inhibitor okadaic acid all dramatically inhibited both PAH induction and H8 potentiation of PAH induction of aldh3 expression but had no effect on induction of cyp1A1 expression in cultured hepatocytes.Both basal and PAH-dependent expression of a chloramphenicol acetyltransferase expression plasmid containing approximately 3.5 kilobase pairs of the 5-flanking region of aldh3 (pALDH3.5CAT) were enhanced 3-4-fold by the PKA inhibitor H8 but not by the PKC inhibitor H7 (>20 M). cAMP analogs, activators of PKA activity, or protein phosphatase inhibitors diminished expression of the reporter gene in a manner identical to the native gene in cultured rat hepatocytes. Using deletion analysis of the pALDH3.5CAT construct, we demonstrated the existence of a negative regulatory region in the 5-flanking region between ؊1057 and ؊991 base pairs which appears to be responsible for the cAMP-dependent regulation of this gene under both basal and PAH-induced conditions. At least two apparently independent mechanisms which involve protein phosphorylation regulate aldh3 expression. One involves function of the Ah receptor which requires PKC protein phosphorylation to positively regulate both aldh3 and cyp1A1 gene expression and the other a cAMP-responsive process which allows PKA activity to negatively regulate expression of aldh3 under either basal or inducible conditions.The aldehyde dehydrogenases (ALDH, 1 aldehyde NAD(P) ϩ oxidoreductase EC 1.2.1.3) are a family of NAD(P) ϩ -dependent enzymes that oxidize a broad class of alde...

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The Same Xenobiotic Response Element Is Required for Constitutive and Inducible Expression of the Mammalian Aldehyde Dehydrogenase-3 Gene

Boesch

Miskimins

et al. 1999

Archives of Biochemistry and Biophysics

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Characterization of the rat Class 3 aldehyde dehydrogenase gene promoter

Cited by 20 publications

References 32 publications

Aldehyde dehydrogenase class 3 expression: Identification of a cornea-preferred gene promoter in transgenic mice

Aldehyde dehydrogenase class 3 expression: Identification of a cornea-preferred gene promoter in transgenic mice

cAMP-dependent Negative Regulation of Rat Aldehyde Dehydrogenase Class 3 Gene Expression

The Same Xenobiotic Response Element Is Required for Constitutive and Inducible Expression of the Mammalian Aldehyde Dehydrogenase-3 Gene

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