scholarly journals Interferon-responsive regulatory elements in the promoter of the human 2',5'-oligo(A) synthetase gene.

1987 ◽  
Vol 7 (12) ◽  
pp. 4498-4504 ◽  
Author(s):  
P Benech ◽  
M Vigneron ◽  
D Peretz ◽  
M Revel ◽  
J Chebath
Keyword(s):  
Dna Sequences ◽  
Regulatory Region ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Base Pairs ◽  
Independent Manner ◽  
E Gene ◽  
Constitutive Enhancer ◽  
Chloramphenicol Acetyltransferase Gene ◽  
Simplex Virus

The interferon (IFN)-activated human 2',5'-oligo(A) synthetase E gene contains 11 RNA starts and lacks TATA and CAAT signals. DNA sequences around the promoter make the expression of the chloramphenicol acetyltransferase gene (CAT) inducible over 20-fold by IFN. A 72-base-pair segment (E-IRS) immediately upstream of the RNA starts was defined as being required for IFN-activated expression of the E-gene promoter-CAT constructs and acts in a position-independent manner. It also confers IFN-activated enhancement to the herpes simplex virus thymidine kinase promoter. On this promoter, the 5' part of the E-IRS functions as a constitutive enhancer, while the last 16 base pairs of the E-IRS is sufficient to give IFN-induced expression. On the E-gene promoter, the constitutive enhancer and the IFN-activated sequence are both needed but can be separated. In addition, promoter competition experiments indicate a third regulatory region which helps to repress expression of the E gene in uninduced cells.

Interferon-responsive regulatory elements in the promoter of the human 2',5'-oligo(A) synthetase gene

1987 ◽  
Vol 7 (12) ◽  
pp. 4498-4504
Author(s):  
P Benech ◽  
M Vigneron ◽  
D Peretz ◽  
M Revel ◽  
J Chebath
Keyword(s):  
Dna Sequences ◽  
Regulatory Region ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Base Pairs ◽  
Independent Manner ◽  
E Gene ◽  
Constitutive Enhancer ◽  
Chloramphenicol Acetyltransferase Gene ◽  
Simplex Virus

The interferon (IFN)-activated human 2',5'-oligo(A) synthetase E gene contains 11 RNA starts and lacks TATA and CAAT signals. DNA sequences around the promoter make the expression of the chloramphenicol acetyltransferase gene (CAT) inducible over 20-fold by IFN. A 72-base-pair segment (E-IRS) immediately upstream of the RNA starts was defined as being required for IFN-activated expression of the E-gene promoter-CAT constructs and acts in a position-independent manner. It also confers IFN-activated enhancement to the herpes simplex virus thymidine kinase promoter. On this promoter, the 5' part of the E-IRS functions as a constitutive enhancer, while the last 16 base pairs of the E-IRS is sufficient to give IFN-induced expression. On the E-gene promoter, the constitutive enhancer and the IFN-activated sequence are both needed but can be separated. In addition, promoter competition experiments indicate a third regulatory region which helps to repress expression of the E gene in uninduced cells.

scholarly journals Interaction between two different regulatory elements activates the murine alpha A-crystallin gene promoter in explanted lens epithelia.

1987 ◽  
Vol 7 (5) ◽  
pp. 1807-1814 ◽  
Author(s):  
A B Chepelinsky ◽  
B Sommer ◽  
J Piatigorsky
Keyword(s):  
Dna Sequences ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Regulatory Sequences ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Promoter Sequences ◽  
Hybrid Genes ◽  
Cat Gene ◽  
Cat Expression

Previous experiments have indicated that 5' flanking DNA sequences (nucleotides-366 to +46) are capable of regulating the lens-specific transcription of the murine alpha A-crystallin gene. Here we have analyzed these 5' regulatory sequences by transfecting explanted embryonic chicken lens epithelia with different alpha A-crystallin-CAT (chloramphenicol acetyltransferase) hybrid genes (alpha A-crystallin promoter sequences fused to the bacterial CAT gene in the pSVO-CAT expression vector). The results indicated the presence of a proximal (-88 to +46) and a distal (-111 to -88) domain which must interact for promoter function. Deletion experiments showed that the sequence between -88 and -60 was essential for function of the proximal domain in the explanted epithelia. A synthetic oligonucleotide containing the sequence between -111 and -84 activated the proximal domain when placed in either orientation 57 base pairs upstream from position -88 of the alpha A-crystallin-CAT hybrid gene.

Interaction between two different regulatory elements activates the murine alpha A-crystallin gene promoter in explanted lens epithelia

1987 ◽  
Vol 7 (5) ◽  
pp. 1807-1814
Author(s):  
A B Chepelinsky ◽  
B Sommer ◽  
J Piatigorsky
Keyword(s):  
Dna Sequences ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Regulatory Sequences ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Promoter Sequences ◽  
Hybrid Genes ◽  
Cat Gene ◽  
Cat Expression

Previous experiments have indicated that 5' flanking DNA sequences (nucleotides-366 to +46) are capable of regulating the lens-specific transcription of the murine alpha A-crystallin gene. Here we have analyzed these 5' regulatory sequences by transfecting explanted embryonic chicken lens epithelia with different alpha A-crystallin-CAT (chloramphenicol acetyltransferase) hybrid genes (alpha A-crystallin promoter sequences fused to the bacterial CAT gene in the pSVO-CAT expression vector). The results indicated the presence of a proximal (-88 to +46) and a distal (-111 to -88) domain which must interact for promoter function. Deletion experiments showed that the sequence between -88 and -60 was essential for function of the proximal domain in the explanted epithelia. A synthetic oligonucleotide containing the sequence between -111 and -84 activated the proximal domain when placed in either orientation 57 base pairs upstream from position -88 of the alpha A-crystallin-CAT hybrid gene.

Distinct regulation of the interleukin-1 and interleukin-6 response elements of the rat haptoglobin gene in rat and human hepatoma cells

1990 ◽  
Vol 10 (11) ◽  
pp. 5967-5976
Author(s):  
H Baumann ◽  
K K Morella ◽  
G P Jahreis ◽  
S Marinković
Keyword(s):  
Interleukin 1 ◽  
Gene Promoter ◽  
Cell Types ◽  
Regulatory Elements ◽  
Hepatoma Cells ◽  
Expression Vectors ◽  
Human Hepatoma ◽  
Human Hepatoma Cells ◽  
Cis Acting ◽  
Chloramphenicol Acetyltransferase Gene

The transcription rate of the haptoglobin (Hp) gene is stimulated by interleukin-1 (IL-1), IL-6, and dexamethasone in rat hepatoma (H-35) cells. To identify the cis-acting regulatory elements responsive to these hormones, various lengths of 5' Hp gene-flanking regions, including the promoter, were inserted into chloramphenicol acetyltransferase gene expression vectors and transiently introduced into H-35 cells. The first 4 kb of 5' region mediated a severalfold increase in expression after treatment with IL-6 and dexamethasone. No response to IL-1 was detectable. When, however, upstream sequences were deleted to position -165 relative to the transcription start site, a significant stimulation by IL-1 was gained without appreciably affecting the IL-6 response. With the apparent removal of an inhibitory sequence, the promoter-proximal 165-bp region also displayed a severalfold enhanced response to the combination of dexamethasone, IL-1, and IL-6. The sequence from -165 to -147, termed the A-element, was found to be crucial for all hormone regulatory functions. Two copies of the A-element linked to a heterologous promoter responded to the three hormones, but to a lesser degree than in the Hp gene promoter context. The regulatory elements of the rat Hp gene were similarly active in human hepatoma cells. Optimal regulation by IL-6 in HepG2 cells was, however, independent of the A-element. The A-element functioned in these cells exclusively as an IL-1 response sequence. The results suggest that genomic sequences upstream of the rat Hp gene suppress the regulation by specific cytokines more prominently in transient expression assays than in the normal chromosomal context. Moreover, the functional comparison indicated that specific regulatory regions of the rat Hp gene do not function identically in different hepatic cell types.

scholarly journals Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter.

1990 ◽  
Vol 10 (7) ◽  
pp. 3834-3837 ◽  
Author(s):  
H H Roehl ◽  
S E Conrad
Keyword(s):  
Thymidine Kinase ◽  
Regulatory Region ◽  
Gene Promoter ◽  
S Phase ◽  
Deletion Mutants ◽  
Thymidine Kinase Gene ◽  
Base Pairs ◽  
Kinase Gene ◽  
Transcriptional Start Sites ◽  
Neomycin Resistance

We have identified a regulatory region in the human thymidine kinase gene promoter. A set of promoter deletion mutants was constructed, linked to the bacterial neomycin resistance gene, and stably transfected into Rat3 cells. It was shown that the region between 135 and 67 base pairs upstream of the cap site is required for conveying G1-S-phase regulation to the linked neo gene. In addition, primer extension assays demonstrated that the same transcriptional start sites were used in G1- and S-phase cells and in the various deletion mutants tested.

Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter

1990 ◽  
Vol 10 (7) ◽  
pp. 3834-3837
Author(s):  
H H Roehl ◽  
S E Conrad
Keyword(s):  
Thymidine Kinase ◽  
Regulatory Region ◽  
Gene Promoter ◽  
S Phase ◽  
Deletion Mutants ◽  
Thymidine Kinase Gene ◽  
Base Pairs ◽  
Kinase Gene ◽  
Transcriptional Start Sites ◽  
Neomycin Resistance

We have identified a regulatory region in the human thymidine kinase gene promoter. A set of promoter deletion mutants was constructed, linked to the bacterial neomycin resistance gene, and stably transfected into Rat3 cells. It was shown that the region between 135 and 67 base pairs upstream of the cap site is required for conveying G1-S-phase regulation to the linked neo gene. In addition, primer extension assays demonstrated that the same transcriptional start sites were used in G1- and S-phase cells and in the various deletion mutants tested.

scholarly journals Regulation of cyclin E gene expression by the human papillomavirus type 16 E7 oncoprotein

1999 ◽  
Vol 80 (8) ◽  
pp. 2103-2113 ◽  
Author(s):  
Beate Vogt ◽  
Karin Zerfaß-Thome ◽  
Almut Schulze ◽  
Jürgen W. Botz ◽  
Werner Zwerschke ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Cyclin E ◽  
Regulatory Region ◽  
Regulatory Elements ◽  
E Gene ◽  
Human Papillomavirus Type 16 ◽  
Activation Of Transcription ◽  
Cellular Transcription

In this study, we characterized the 5′ regulatory region of the murine cyclin E gene and analysed activation of the gene by the E7 oncogene of human papillomavirus type 16 in transfection experiments. We found that the murine cyclin E promoter is composed of multiple regulatory elements, and we present evidence for at least two independent transcription units, designated P1 and P2. Overlapping binding sites for the cellular transcription factors Sp1 and E2F were identified in both promoters, and we found that E2F-mediated activation of transcription is inhibited by Sp1 in cotransfection experiments. The E2F/Sp1 binding sites contribute to transcriptional activation by E7, and the data suggest that the cyclin E gene is rendered E7-inducible through the combination of several cis-acting elements which display only weak intrinsic responsiveness to E7.

Regulatory elements mediating transcription from the Drosophila melanogaster actin 5C proximal promoter

1990 ◽  
Vol 10 (1) ◽  
pp. 206-216
Author(s):  
Y T Chung ◽  
E B Keller
Keyword(s):  
Drosophila Melanogaster ◽  
Transient Expression ◽  
Promoter Activity ◽  
Regulatory Element ◽  
Regulatory Elements ◽  
Proximal Promoter ◽  
Base Pairs ◽  
Chloramphenicol Acetyltransferase Gene ◽  
Constitutive Synthesis

The major cytoskeletal actin gene of Drosophila melanogaster, the actin 5C gene, has two promoters, the proximal one of which controls constitutive synthesis of actin in all growing tissues. To locate regulatory elements required for constitutive activity of the proximal promoter, mutants of this promoter were fused to the bacterial chloramphenicol acetyltransferase gene and assayed for transient expression activity in cultured Drosophila embryonic Schneider line 2 cells. An essential regulatory element has been located 313 base pairs upstream from the cap site. Deletion of this element lowered expression to one-third of the wild-type level. The element has the sequence AAGTTGTAGTTG, as shown by protein-binding footprinting with the reagent methidiumpropyl-EDTA-Fe(II). This element is probably not a general one, since it was not detected in a search of the published 5'-flanking sequences of 27 Drosophila genes. In addition to this regulatory element, there are five GAGA elements in the actin 5C proximal promoter, some or all of which are essential for the promoter activity as shown by an in vivo competition assay. Although this promoter has no classical TATA element, there is an essential promoter region about 35 base pairs upstream from the cap site that could be a TATA surrogate. The promoter also shows sequences homologous to the alcohol dehydrogenase factor 1-binding site and to the core of the vertebrate serum response element, but mutations of these sites did not affect promoter activity in transient expression assays.

scholarly journals The activity of the highly inducible mouse phenylalanine hydroxylase gene promoter is dependent upon a tissue-specific, hormone-inducible enhancer.

1996 ◽  
Vol 16 (6) ◽  
pp. 3125-3137 ◽  
Author(s):  
D M Faust ◽  
A M Catherin ◽  
S Barbaux ◽  
L Belkadi ◽  
T Imaizumi-Scherrer ◽  
...  
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Phenylalanine Hydroxylase ◽  
Regulatory Region ◽  
Gene Promoter ◽  
Inducible Promoter ◽  
Regulatory Elements ◽  
Tyrosine Aminotransferase ◽  
Tissue Specific ◽  
Hydroxylase Gene ◽  
Phenylalanine Hydroxylase Gene

Expression of the phenylalanine hydroxylase gene in livers and kidneys of rodents is activated at birth and is induced by glucocorticoids and cyclic AMP in the liver. Regulatory elements in a 10-kb fragment upstream of the mouse gene have been characterized. The promoter lacks TAATA and CCAAT consensus sequences and shows only extremely weak activity in transitory expression assays with phenylalanine hydroxylase-producing hepatoma cells. No key elements for regulation of promoter activity are localized within 2 kb of upstream sequences. However, a liver-specific DNase I-hypersensitive site at kb -3.5 comprises a tissue-specific and hormone-inducible enhancer. This enhancer contains multiple protein binding sites, including sites for ubiquitous factors (NF1 and AP1), the glucocorticoid receptor, and the hepatocyte-enriched transcription factors hepatocyte nuclear factor 1 (HNF1) and C/EBP. Mutation revealed that the last two sites are critical not only for basal activity but also for obtaining a maximal hormone response. Efficient transcription from the highly inducible promoter shows absolute dependence upon the enhancer at kb - 3.5, which in turn requires HNF1 and C/EBP as well as hormones. The regulatory region of the mouse phenylalanine hydroxylase gene differs totally from that of humans, even though the genes of both species are expressed essentially in the liver. Furthermore, the phenylalanine hydroxylase gene of mice shows an expression pattern very similar to those of the rodent tyrosine aminotransferase and phosphoenolpyruvate carboxykinase genes, yet each shows a different organization of its regulatory region.

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