scholarly journals cis-acting translational effects of the 5' noncoding region of c-myc mRNA.

1988 ◽  
Vol 8 (7) ◽  
pp. 2875-2883 ◽  
Author(s):  
N Parkin ◽  
A Darveau ◽  
R Nicholson ◽  
N Sonenberg
Keyword(s):  
Inhibitory Effect ◽  
Noncoding Region ◽  
Translational Efficiency ◽  
Cis Acting ◽  
Cell Extracts ◽  
Negative Effect ◽  
Reticulocyte Lysate ◽  
Established Cell

We have previously shown that the 5' noncoding region of mouse c-myc mRNA has a negative effect on translational efficiency in a rabbit reticulocyte lysate (A. Darveau, J. Pelletier, and N. Sonenberg, Proc. Natl. Acad. Sci. USA 82:2315-2319, 1985). We wanted to localize and characterize the inhibitory translational element(s) in the mRNA and to study its effect in other in vitro and in vivo systems. Here we report that the restrictive element is confined to a 240-nucleotide sequence of the 5' noncoding region of mouse c-myc mRNA and that this sequence acts in cis to inhibit the translation of a heterologous mRNA. In addition, we report that the cis-inhibitory effect is also exhibited in microinjected Xenopus oocytes and wheat-germ extracts but not in HeLa cell extracts. Transfection of corresponding plasmid DNA constructs into several established cell lines did not produce the cis-inhibitory effect. A model to explain these results is presented.

cis-acting translational effects of the 5' noncoding region of c-myc mRNA

1988 ◽  
Vol 8 (7) ◽  
pp. 2875-2883
Author(s):  
N Parkin ◽  
A Darveau ◽  
R Nicholson ◽  
N Sonenberg
Keyword(s):  
Inhibitory Effect ◽  
Noncoding Region ◽  
Translational Efficiency ◽  
Cis Acting ◽  
Cell Extracts ◽  
Negative Effect ◽  
Reticulocyte Lysate ◽  
Established Cell

We have previously shown that the 5' noncoding region of mouse c-myc mRNA has a negative effect on translational efficiency in a rabbit reticulocyte lysate (A. Darveau, J. Pelletier, and N. Sonenberg, Proc. Natl. Acad. Sci. USA 82:2315-2319, 1985). We wanted to localize and characterize the inhibitory translational element(s) in the mRNA and to study its effect in other in vitro and in vivo systems. Here we report that the restrictive element is confined to a 240-nucleotide sequence of the 5' noncoding region of mouse c-myc mRNA and that this sequence acts in cis to inhibit the translation of a heterologous mRNA. In addition, we report that the cis-inhibitory effect is also exhibited in microinjected Xenopus oocytes and wheat-germ extracts but not in HeLa cell extracts. Transfection of corresponding plasmid DNA constructs into several established cell lines did not produce the cis-inhibitory effect. A model to explain these results is presented.

cis- and trans-acting regulatory elements of the yeast URA3 promoter

1990 ◽  
Vol 10 (10) ◽  
pp. 5257-5270
Author(s):  
A Roy ◽  
F Exinger ◽  
R Losson
Keyword(s):  
Specific Binding ◽  
Point Mutations ◽  
Regulatory Elements ◽  
Cis Acting ◽  
Cell Extracts ◽  
Dnase I Footprinting ◽  
Pair Sequence ◽  
Dyad Symmetry

Expression of the yeast pyrimidine biosynthetic gene, URA3, is induced three- to fivefold in response to uracil starvation, and this regulation is mediated by the transcriptional activator PPR1 (pyrimidine pathway regulator 1). In this study, we have analyzed the regulatory elements of the URA3 promoter by DNase I footprinting, using partially purified yeast cell extracts, by deletion mutagenesis, and by 5'-end mapping of RNA transcripts. Two DNA-binding activities have been detected, and at least four distinct cis-acting regions have been identified. A region rich in poly(dA-dT) serves as an upstream promoter element necessary for the basal level of URA3 expression. A 16-base-pair sequence with dyad symmetry acts acts as a uracil-controlled upstream activating site (UASURA) and shows a specific binding only with cell extracts from strains overproducing PPR1. This in vitro binding does not require dihydroorotic acid, the physiological inducer of URA3. The TATA region appears to be composed of two functionally distinct (constitutive and regulatory) elements. Two G + A-rich regions surrounding this TATA box bind an unidentified factor called GA-binding factor. The 5' copy, GA1, is involved in PPR1 induction and overlaps the constitutive TATA region. The 3' region, GA2, is necessary for maximal expression. Neither of these GA sequences acts as a UAS in a CYC1-lacZ context. The promoters of the unlinked but coordinately regulated URA1 and URA4 genes contain highly conserved copies of the UASURA sequence, which prompted us to investigate the effects of many point mutations within this UASURA sequence on PPR1-dependent binding. In this way, we have identified the most important residues of this binding site and found that a nonsymmetrical change of these bases is sufficient to prevent the specific binding and to suppress the UASURA activity in vivo. In addition, we showed that UASURA contains a constitutive activating element which can stimulate transcription from a heterologous promoter independently of dihydroorotic acid and PPR1.

scholarly journals cis- and trans-acting regulatory elements of the yeast URA3 promoter.

1990 ◽  
Vol 10 (10) ◽  
pp. 5257-5270 ◽  
Author(s):  
A Roy ◽  
F Exinger ◽  
R Losson
Keyword(s):  
Specific Binding ◽  
Point Mutations ◽  
Regulatory Elements ◽  
Cis Acting ◽  
Cell Extracts ◽  
Dnase I Footprinting ◽  
Pair Sequence ◽  
Dyad Symmetry

Expression of the yeast pyrimidine biosynthetic gene, URA3, is induced three- to fivefold in response to uracil starvation, and this regulation is mediated by the transcriptional activator PPR1 (pyrimidine pathway regulator 1). In this study, we have analyzed the regulatory elements of the URA3 promoter by DNase I footprinting, using partially purified yeast cell extracts, by deletion mutagenesis, and by 5'-end mapping of RNA transcripts. Two DNA-binding activities have been detected, and at least four distinct cis-acting regions have been identified. A region rich in poly(dA-dT) serves as an upstream promoter element necessary for the basal level of URA3 expression. A 16-base-pair sequence with dyad symmetry acts acts as a uracil-controlled upstream activating site (UASURA) and shows a specific binding only with cell extracts from strains overproducing PPR1. This in vitro binding does not require dihydroorotic acid, the physiological inducer of URA3. The TATA region appears to be composed of two functionally distinct (constitutive and regulatory) elements. Two G + A-rich regions surrounding this TATA box bind an unidentified factor called GA-binding factor. The 5' copy, GA1, is involved in PPR1 induction and overlaps the constitutive TATA region. The 3' region, GA2, is necessary for maximal expression. Neither of these GA sequences acts as a UAS in a CYC1-lacZ context. The promoters of the unlinked but coordinately regulated URA1 and URA4 genes contain highly conserved copies of the UASURA sequence, which prompted us to investigate the effects of many point mutations within this UASURA sequence on PPR1-dependent binding. In this way, we have identified the most important residues of this binding site and found that a nonsymmetrical change of these bases is sufficient to prevent the specific binding and to suppress the UASURA activity in vivo. In addition, we showed that UASURA contains a constitutive activating element which can stimulate transcription from a heterologous promoter independently of dihydroorotic acid and PPR1.

Regulation of in vitro and in vivo transcription of early-region IV of adenovirus type 5 by multiple cis-acting elements

1987 ◽  
Vol 7 (7) ◽  
pp. 2578-2587
Author(s):  
S Hanaka ◽  
T Nishigaki ◽  
P A Sharp ◽  
H Handa
Keyword(s):  
Transcriptional Activity ◽  
Adenovirus Type ◽  
Tata Box ◽  
Upstream Region ◽  
Inverted Repeats ◽  
Cis Acting ◽  
Early Region ◽  
Cell Extracts

A series of deletion mutants spanning the promoter of the adenovirus early-region IV (EIV) gene were tested for transcriptional activity, using both in vitro and in vivo assays. Four distinct domains had additive effects on efficient transcription from the EIV promoter in HeLa whole-cell extracts. The first resided 20 to 27 bases upstream of the initiation site and included the TATA box. Deletion of the TATA box drastically reduced the transcriptional activity in vitro but had a lesser effect in vivo. The second region extended from -32 to -177 and contained two 17-base-pair inverted repeats, centered around -40 and -162. Sequences lying between -140 and -173 were important for efficient transcription since deletion of this region reduced the activity fourfold. Deletion of either one of the two inverted repeats or insertion of DNA fragments between them resulted in the synthesis of extra transcripts that initiated at sites upstream from the EIV site. The third region was located between -198 and -250 and contains three guanosine-plus-cytosine-rich sequences, present around -212 (GGGCGG), -233 (GGGCGG), and -251 (CGCGGG). The fourth, most upstream region was located between -260 and -307. Deletion of this region, which contains the NF-1 factor-binding site, slightly reduced transcriptional activity both in vivo and in vitro. The data indicate that multiple cis-acting elements are required for efficient transcription from the EIV promoter in both in vitro and in vivo systems.

The 5' noncoding region of the human leukemia-associated oncogene BCR/ABL is a potent inhibitor of in vitro translation

1989 ◽  
Vol 9 (11) ◽  
pp. 5234-5238
Author(s):  
A J Muller ◽  
O N Witte
Keyword(s):  
Philadelphia Chromosome ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Noncoding Region ◽  
Myelogenous Leukemia ◽  
In Vitro Translation ◽  
Translational Efficiency ◽  
Human Leukemia ◽  
Untranslated Sequence

The mRNA encoding the chimeric BCR/ABL oncogene, which is transcribed from the Philadelphia chromosome in human chronic myelogenous leukemia, has a 5' noncoding sequence greater than 500 bases in length which is highly GC rich and contains a short open reading frame. This untranslated sequence has a dramatic inhibitory effect upon translational efficiency in vitro. However, when BCR/ABL message is expressed in certain cell types such as the NIH 3T3 cell line, the 5' noncoding region has little inhibitory effect on translational efficiency.

FXR-activating ligands inhibit rabbit ASBT expression via FXR-SHP-FTF cascade

2005 ◽  
Vol 288 (1) ◽  
pp. G60-G66 ◽  
Author(s):  
Hai Li ◽  
Frank Chen ◽  
Quan Shang ◽  
Luxing Pan ◽  
Benjamin L. Shneider ◽  
...  
Keyword(s):  
Bile Acid ◽  
Binding Site ◽  
Bile Acids ◽  
Promoter Activity ◽  
Deoxycholic Acid ◽  
Inhibitory Effect ◽  
Cis Acting ◽  
Regulatory Cascade

The regulation of the rabbit apical sodium-dependent bile acid transporter (ASBT) was studied both in vivo and in vitro. New Zealand White rabbits were fed 0.5% deoxycholic acid (DCA) or SC-435, a competitive ASBT inhibitor, for 1 wk. In DCA-fed rabbits, ASBT expression was repressed, associated with activated FXR, and evidenced by increased ileal short heterodimer partner (SHP) mRNA. Feeding SC-435 to the rabbits blocked bile acid absorption, decreased SHP mRNA, and increased ASBT expression. A 1.9-kb rabbit ASBT 5′-flanking region (promoter) was cloned, and a cis-acting element for α-fetoprotein transcription factor (FTF) was identified (−1166/ −1158). The effects of transcriptional factors and different bile acids on the rabbit ASBT promoter were studied in Caco-2 cells. FTF stimulated the rabbit ASBT promoter activity fourfold but not after the FTF binding site was deleted from the promoter. Increasing the SHP protein notably inhibited FTF-dependent trans-activation of rabbit ASBT. Adding hydrophobic bile acids deoxycholic acid, chenodeoxycholic acid, and cholic acid, activating ligands for FXR, inhibited rabbit ASBT promoter activity in Caco-2 cells, but this inhibitory effect was abolished after the FTF binding site was deleted. Ursodeoxycholic acid and ursocholic acid, nonactivating ligands for FXR, did not repress ASBT promoter activity. Thus the rabbit ASBT promoter is negative-feedback regulated by bile acids via a functional FTF binding site. Only FXR-activating ligands can downregulate rabbit ASBT expression through the regulatory cascade FXR-SHP-FTF.

scholarly journals Regulation of in vitro and in vivo transcription of early-region IV of adenovirus type 5 by multiple cis-acting elements.

1987 ◽  
Vol 7 (7) ◽  
pp. 2578-2587 ◽  
Author(s):  
S Hanaka ◽  
T Nishigaki ◽  
P A Sharp ◽  
H Handa
Keyword(s):  
Transcriptional Activity ◽  
Adenovirus Type ◽  
Tata Box ◽  
Upstream Region ◽  
Inverted Repeats ◽  
Cis Acting ◽  
Early Region ◽  
Cell Extracts

A series of deletion mutants spanning the promoter of the adenovirus early-region IV (EIV) gene were tested for transcriptional activity, using both in vitro and in vivo assays. Four distinct domains had additive effects on efficient transcription from the EIV promoter in HeLa whole-cell extracts. The first resided 20 to 27 bases upstream of the initiation site and included the TATA box. Deletion of the TATA box drastically reduced the transcriptional activity in vitro but had a lesser effect in vivo. The second region extended from -32 to -177 and contained two 17-base-pair inverted repeats, centered around -40 and -162. Sequences lying between -140 and -173 were important for efficient transcription since deletion of this region reduced the activity fourfold. Deletion of either one of the two inverted repeats or insertion of DNA fragments between them resulted in the synthesis of extra transcripts that initiated at sites upstream from the EIV site. The third region was located between -198 and -250 and contains three guanosine-plus-cytosine-rich sequences, present around -212 (GGGCGG), -233 (GGGCGG), and -251 (CGCGGG). The fourth, most upstream region was located between -260 and -307. Deletion of this region, which contains the NF-1 factor-binding site, slightly reduced transcriptional activity both in vivo and in vitro. The data indicate that multiple cis-acting elements are required for efficient transcription from the EIV promoter in both in vitro and in vivo systems.

scholarly journals The 5' noncoding region of the human leukemia-associated oncogene BCR/ABL is a potent inhibitor of in vitro translation.

1989 ◽  
Vol 9 (11) ◽  
pp. 5234-5238 ◽  
Author(s):  
A J Muller ◽  
O N Witte
Keyword(s):  
Philadelphia Chromosome ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Noncoding Region ◽  
Myelogenous Leukemia ◽  
In Vitro Translation ◽  
Translational Efficiency ◽  
Human Leukemia ◽  
Untranslated Sequence

The mRNA encoding the chimeric BCR/ABL oncogene, which is transcribed from the Philadelphia chromosome in human chronic myelogenous leukemia, has a 5' noncoding sequence greater than 500 bases in length which is highly GC rich and contains a short open reading frame. This untranslated sequence has a dramatic inhibitory effect upon translational efficiency in vitro. However, when BCR/ABL message is expressed in certain cell types such as the NIH 3T3 cell line, the 5' noncoding region has little inhibitory effect on translational efficiency.

The Effect of Barbituric Acid Derivatives on Platelet Function in Vitro and in Vivo

1973 ◽  
Vol 30 (02) ◽  
pp. 315-326
Author(s):  
J. Heinz Joist ◽  
Jean-Pierre Cazenave ◽  
J. Fraser Mustard
Keyword(s):  
Platelet Aggregation ◽  
Platelet Function ◽  
Barbituric Acid ◽  
Platelet Rich Plasma ◽  
Inhibitory Effect ◽  
Primary Response ◽  
Sodium Pentobarbital ◽  
Barbituric Acid Derivatives

SummarySodium pentobarbital (SPB) and three other barbituric acid derivatives were found to inhibit platelet function in vitro. SPB had no effect on the primary response to ADP of platelets in platelet-rich plasma (PRP) or washed platelets but inhibited secondary aggregation induced by ADP in human PRP. The drug inhibited both phases of aggregation induced by epinephrine. SPB suppressed aggregation and the release reaction induced by collagen or low concentrations of thrombin, and platelet adherence to collagen-coated glass tubes. The inhibition by SPB of platelet aggregation was readily reversible and isotopically labeled SPB did not become firmly bound to platelets. No inhibitory effect on platelet aggregation induced by ADP, collagen, or thrombin could be detected in PRP obtained from rabbits after induction of SPB-anesthesia.

Inhibition of Human and Animal Platelet Adhesiveness to Glass Bead Columns by Adenosine, Dipyridamole, Chlorpromazine and Acetylsalicylic Acid

1976 ◽  
Vol 36 (02) ◽  
pp. 401-410 ◽  
Author(s):  
Buichi Fujttani ◽  
Toshimichi Tsuboi ◽  
Kazuko Takeno ◽  
Kouichi Yoshida ◽  
Masanao Shimizu
Keyword(s):  
Guinea Pig ◽  
Acetylsalicylic Acid ◽  
Guinea Pigs ◽  
Glass Bead ◽  
Animal Species ◽  
Inhibitory Effect ◽  
Rabbit Platelet ◽  
Platelet Adhesiveness

SummaryThe differences among human, rabbit and guinea-pig platelet adhesiveness as for inhibitions by adenosine, dipyridamole, chlorpromazine and acetylsalicylic acid are described, and the influence of measurement conditions on platelet adhesiveness is also reported. Platelet adhesiveness of human and animal species decreased with an increase of heparin concentrations and an increase of flow rate of blood passing through a glass bead column. Human and rabbit platelet adhesiveness was inhibited in vitro by adenosine, dipyridamole and chlorpromazine, but not by acetylsalicylic acid. On the other hand, guinea-pig platelet adhesiveness was inhibited by the four drugs including acetylsalicylic acid. In in vivo study, adenosine, dipyridamole and chlorpromazine inhibited platelet adhesiveness in rabbits and guinea-pigs. Acetylsalicylic acid showed the inhibitory effect in guinea-pigs, but not in rabbits.

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