Studies on the Structure of Genes Expressed during Development

Sim, Gek-Kee; Efstratiadis, Argiris; Jones, Cynthia; Kafatos, Fotis C.; Köehler, Michael; Kronenberg, Henry M.; Maniatis, Tom; Regier, Jerome C.; Roberts, Brett; Rosenthal, Noël F.

doi:10.1101/sqb.1978.042.01.095

Cited by 9 publications

(7 citation statements)

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“…Cycloheximide also superinduced ferredoxin expression when added together with 8-Br-cAMP. It indicated that new protein synthesis is not required in Yl cells, although stimulation of ferredoxin RNA by cAMP is a late Sim et al (1978) aAmino acid #14 to amino acid #21 of the coding sequence of the human GAP gene. bThe transcription start site is defined as +1.…”

Section: Regulation Offerredoxin Gene Expressionmentioning

confidence: 97%

Differential Regulation of theCYP11A1(P450scc) and Ferredoxin Genes in Adrenal and Placental Cells

Guo¹,

Huang²,

Chung³

1993

DNA and Cell Biology

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The regulation of the genes encoding cholesterol side-chain cleavage enzyme (P450scc) and ferredoxin, two components in the first step of steroid synthetic pathways, was studied by RNA analyses of endogenous and transfected genes. cAMP rather than calcium was the major secondary messenger that stimulated expression of both P450scc and ferredoxin genes in human placental JEG-3 cells. The effect of cAMP on P450scc expression was abolished by cycloheximide in JEG-3 cells, but it was superinduced in mouse adrenal Y1 cells. For ferredoxin expression, both reagents have synergistic effect in Y1 and JEG-3 cells. To test the mechanism of regulation, DNA segments containing regulatory elements of the P450scc and ferredoxin genes were connected to reporter genes and analyzed in cotransfection experiments. The results showed that the proximal cAMP-responsive sequences of both P450scc and ferredoxin genes were stimulated by cAMP early in both Y1 and JEG-3 cells, requiring no new protein synthesis. This indicates a common mechanism for the regulated expression of both genes. P450scc possessed an additional upstream cAMP-responsive sequence that also responded to cAMP induction in a different manner from the proximal element. The presence of additional upstream regulatory elements makes it possible for the P450scc gene to be further regulated.

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Section: Regulation Offerredoxin Gene Expressionmentioning

confidence: 97%

Differential Regulation of theCYP11A1(P450scc) and Ferredoxin Genes in Adrenal and Placental Cells

Guo¹,

Huang²,

Chung³

1993

DNA and Cell Biology

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“…The NheI site is at position k738 in the pig APN gene [22], the NcoI site is at position j166 in the rabbit β-globin gene [25] and 46 nucleotides are derived from the pSP72 polylinker.…”

Section: Template For In Vitro Transcription Of the Globin Antisense mentioning

confidence: 99%

An enhancer with cell-type dependent activity is located between the myeloid and epithelial aminopeptidase N (CD 13) promoters

Olsen

Kokholm

Troelsen

et al. 1997

Biochemical Journal

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The 5h flanking region of the gene encoding the small intestinal brush-border peptidase aminopeptidase N (APN) was screened for the presence of enhancer regions. A 300 bp region with enhancer activity was identified 2.7 kb upstream of the transcriptional start site which is used in epithelial cells. The enhancer stimulated transcription from a heterologous promoter (the simian virus 40 early promoter) in a position-and orientationindependent manner. The activity of the enhancer is cell-type dependent and it is active in liver (HepG2), intestinal (Caco-2) and myeloid (K562) cells. As the epithelial APN promoter is active in the first two cell-types and the myeloid APN promoter in the last, the results may suggest that the enhancer, through

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“…Because the hybrid plasmid PflG1 has been shown to contain the complete coding sequence for rabbit globin (19,20), both the total linearized recombinant plasmid and the purified globin sequence thereof were used to demonstrate the application of the HART method as a general approach to the definition of structural gene sequences within recombinant DNAs.…”

mentioning

confidence: 99%

Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.

Paterson¹,

Roberts²,

Kuff³

1977

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

458

163

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We present a simple method for directly correlating structural gene sequences in DNA with their corresponding mRNAs. This is based upon the fact that mRNA hybridized with its complementary DNA will not direct the cellfree synthesis of a complete polypeptide. Full translational activity of the mRNA is recovered upon the heat melting of the hybrid. Utilizing the rabbit P globin clone PPG1, we demonstrate the application of hybrid-arrested translation for the identification of structural gene sequences within recombinant DNA molecules. In addition, the method is used to locate and order precisely several adenovirus 2 polypeptides within the viral genome. We have developed a method with which to analyze the relationship between DNA sequences, their corresponding messenger RNAs, and the proteins for which they code. This approach is based upon the observation that mRNA in hybrid form with its complementary DNA (cDNA) is not translated in eukaryotic cell-free systems, while heat dissociation of the hybrid reinstates complete translational activity (1). This observation has been extended to double-stranded DNAs by carrying out the hybridization reactions in a concentration of formamide that favors the formation of DNA-RNA hybrids but essentially prevents DNA-DNA reannealing (2)(3)(4)

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Studies on the Structure of Genes Expressed during Development

Cited by 9 publications

References 0 publications

Differential Regulation of theCYP11A1(P450scc) and Ferredoxin Genes in Adrenal and Placental Cells

Differential Regulation of theCYP11A1(P450scc) and Ferredoxin Genes in Adrenal and Placental Cells

An enhancer with cell-type dependent activity is located between the myeloid and epithelial aminopeptidase N (CD 13) promoters

Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.

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