scholarly journals Determinants of rat albumin promoter tissue specificity analyzed by an improved transient expression system.

1987 ◽  
Vol 7 (7) ◽  
pp. 2425-2434 ◽  
Author(s):  
J M Heard ◽  
P Herbomel ◽  
M O Ott ◽  
A Mottura-Rollier ◽  
M Weiss ◽  
...  
Keyword(s):  
Transient Expression ◽  
Tissue Specificity ◽  
Transcriptional Start Site ◽  
Expression System ◽  
Hepatoma Cells ◽  
Sequence Motif ◽  
Specific Expression ◽  
Tissue Specific ◽  
Albumin Gene ◽  
Ccaat Box

The 150-base-pairs region located upstream of the transcriptional start site of the rat albumin gene contains all of the critical sequences necessary for this gene's tissue-specific expression in rat hepatoma cells. In transient expression assays using an improved CAT system or direct mRNA analysis we were able to detect a faithful transcription from the albumin promoter in albumin-negative dedifferentiated H5 hepatoma cells which was 250-fold weaker than in differentiated H4II hepatoma cells producing albumin. This strong tissue specificity could be completely overcome through the cis action of a non-tissue-specific enhancer. Two upstream regions from nucleotides -151 to -119 and from -118 to -94, were required for efficient transcription in H4II cells. Each region contained a sequence motif highly conserved among different species. The effect of the -151/-119 region was strictly tissue specific, while the -118/-94 region was also involved in the low level of transcription observed in H5 cells. Finally, sequences between the CCAAT box and the TATA box also contributed to the overall tissue specificity of rat albumin gene transcription.

Determinants of rat albumin promoter tissue specificity analyzed by an improved transient expression system

1987 ◽  
Vol 7 (7) ◽  
pp. 2425-2434
Author(s):  
J M Heard ◽  
P Herbomel ◽  
M O Ott ◽  
A Mottura-Rollier ◽  
M Weiss ◽  
...  
Keyword(s):  
Transient Expression ◽  
Tissue Specificity ◽  
Transcriptional Start Site ◽  
Expression System ◽  
Hepatoma Cells ◽  
Sequence Motif ◽  
Specific Expression ◽  
Tissue Specific ◽  
Albumin Gene ◽  
Ccaat Box

The 150-base-pairs region located upstream of the transcriptional start site of the rat albumin gene contains all of the critical sequences necessary for this gene's tissue-specific expression in rat hepatoma cells. In transient expression assays using an improved CAT system or direct mRNA analysis we were able to detect a faithful transcription from the albumin promoter in albumin-negative dedifferentiated H5 hepatoma cells which was 250-fold weaker than in differentiated H4II hepatoma cells producing albumin. This strong tissue specificity could be completely overcome through the cis action of a non-tissue-specific enhancer. Two upstream regions from nucleotides -151 to -119 and from -118 to -94, were required for efficient transcription in H4II cells. Each region contained a sequence motif highly conserved among different species. The effect of the -151/-119 region was strictly tissue specific, while the -118/-94 region was also involved in the low level of transcription observed in H5 cells. Finally, sequences between the CCAAT box and the TATA box also contributed to the overall tissue specificity of rat albumin gene transcription.

Cooperativity of sequence elements mediates tissue specificity of the rat insulin II gene

1990 ◽  
Vol 10 (4) ◽  
pp. 1784-1788
Author(s):  
Y P Hwung ◽  
Y Z Gu ◽  
M J Tsai
Keyword(s):  
Beta Cell ◽  
Tissue Specificity ◽  
Promoter Element ◽  
Heterologous Promoter ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Insulin Promoter ◽  
Sequence Elements ◽  
Flanking Region

The 5'-flanking region of the rat insulin II gene (-448 to +50) is sufficient for tissue-specific expression. To further determine the tissue-specific cis-acting element(s), important sequences defined by linker-scanning mutagenesis were placed upstream of a heterologous promoter and transfected into insulin-producing and -nonproducing cells. Rat insulin promoter element 3 (RIPE3), which spans from -125 to -86, was shown to confer beta-cell-specific expression in either orientation. However, two subregions of RIPE3, RIPE3a and RIPE3b (defined by linker-scanning mutations), displayed only marginal activities. These results suggest that the two subregions cooperate to confer tissue specificity, presumably via their cognate binding factors.

scholarly journals Development of cDNA normalization system and preliminary transcription analysis of KCS genes in apple tissues

2011 ◽  
Vol 59 (3) ◽  
pp. 9-12 ◽  
Author(s):  
Zsolt Albert ◽  
Cs. Deák ◽  
A. Miskó ◽  
M. Tóth ◽  
I. Papp
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Expression Patterns ◽  
Housekeeping Genes ◽  
Apple Fruit ◽  
Rt Pcr ◽  
Specific Primers ◽  
Specific Expression ◽  
Transcription Analysis ◽  
Tissue Specific

Wax production is an important aspect of apple (Malus domestica Borkh.) fruit development from both theoretical and practical point of views. The complex molecular mechanism that controls wax biosynthesis is still widely unknown but many studies focused on this topic. We aimed to develop further the experimental framework of these efforts with a description of an improved reference genes expression system. Results in the literature show that similarities exist among the expression of some housekeeping genes of different plant species. Based on these considerations and on gene expression data from Arabidopsis thaliana, some genes in apple were assigned for analysis. EST sequences of apple were used to design specific primers for RT-PCR experiments. Isolation of intact RNA from different apple tissues and performing RT-PCR reaction were also key point in obtaining expression patterns. To monitor DNA contamination of the RNA samples, specific primers were used that amplify intron-containing sequences from the cDNA. We found that actin primers can be used for the detection of intron containing genomic DNA, and tubulin primers are good internal controls in RT-PCR experiments. We were able to make a difference between tissue-specific and tissue-independent gene-expression, furthermore we found tissue specific differences between the expression patterns of candidate genes, that are potentially involved in wax-biosynthesis. Our results show that KCS1 and KCS4 are overexpressed in the skin tissue, this could mean that these genes have skin-specific expression in apple fruit.

Tissue-specific expression is conferred by a sequence from the 5′ end of the rat albumin gene.

1984 ◽  
Vol 3 (11) ◽  
pp. 2505-2510 ◽  
Author(s):  
M.O. Ott ◽  
L. Sperling ◽  
P. Herbomel ◽  
M. Yaniv ◽  
M.C. Weiss
Keyword(s):  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Albumin Gene

scholarly journals Cooperativity of sequence elements mediates tissue specificity of the rat insulin II gene.

1990 ◽  
Vol 10 (4) ◽  
pp. 1784-1788 ◽  
Author(s):  
Y P Hwung ◽  
Y Z Gu ◽  
M J Tsai
Keyword(s):  
Beta Cell ◽  
Tissue Specificity ◽  
Promoter Element ◽  
Heterologous Promoter ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Insulin Promoter ◽  
Sequence Elements ◽  
Flanking Region

The 5'-flanking region of the rat insulin II gene (-448 to +50) is sufficient for tissue-specific expression. To further determine the tissue-specific cis-acting element(s), important sequences defined by linker-scanning mutagenesis were placed upstream of a heterologous promoter and transfected into insulin-producing and -nonproducing cells. Rat insulin promoter element 3 (RIPE3), which spans from -125 to -86, was shown to confer beta-cell-specific expression in either orientation. However, two subregions of RIPE3, RIPE3a and RIPE3b (defined by linker-scanning mutations), displayed only marginal activities. These results suggest that the two subregions cooperate to confer tissue specificity, presumably via their cognate binding factors.

Functionally distinct elements are required for expression of the AMPD1 gene in myocytes

1993 ◽  
Vol 13 (9) ◽  
pp. 5854-5860
Author(s):  
T Morisaki ◽  
E W Holmes
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Transcriptional Start Site ◽  
Protein S ◽  
Muscle Fiber Types ◽  
Binding Motif ◽  
Fiber Types ◽  
Specific Expression ◽  
Tissue Specific ◽  
Ampd1 Gene

AMP deaminase (AMPD) is an enzyme found in all eukaryotic cells. Tissue-specific and stage-specific isoforms of this enzyme are found in vertebrates, and expression of these different isoforms is determined by selective expression of the multiple genes. The AMPD1 gene is expressed predominantly in skeletal muscle, in which transcript abundance is controlled by stage-specific and fiber type-specific signals. This enzyme activity is presumed to be important in skeletal muscle because a metabolic myopathy develops in individuals with an inherited deficiency of AMPD1. In the present study, cis- and trans-acting factors that control expression of AMPD1 have been identified. Two cis-acting elements located within 100 nucleotides of the transcriptional start site are required for muscle-specific expression of AMPD1. One element (-100 to -79) behaves like a tissue-specific enhancer, and it interacts with protein(s) found predominantly in nuclei of myoblasts and myotubes. This element is similar in sequence to an MEF2 binding motif, and it contains an A/T core that is essential for enhancer activity and binding of a nuclear protein(s). The second element (-60 to -40) has properties of a stage-specific promoter in that it is essential for muscle-specific expression of the AMPD1 promoter, does not confer muscle-specific expression on a heterologous promoter construct, and interacts with a protein(s) restricted to nuclei of differentiated myotubes. Interaction between these functionally distinct elements may be required for regulating the expression of AMPD1 during myocyte differentiation and in different muscle fiber types.

scholarly journals Cloning and tissue-specific expression of the gene for mouse C-reactive protein

1993 ◽  
Vol 295 (2) ◽  
pp. 379-386 ◽  
Author(s):  
N O Ku ◽  
R F Mortensen
Keyword(s):  
Acute Phase ◽  
Transcriptional Start Site ◽  
Interleukin 1 ◽  
Acute Phase Reactant ◽  
C Reactive Protein ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Reactive Protein ◽  
Phase Reactant

C-reactive protein is a serum acute-phase reactant that increases several thousand-fold in concentration during inflammation in most mammals. However, mouse C-reactive protein is considered to be a minor acute-phase reactant, since its blood level increases only from approx. 0.1 to 1-2 micrograms/ml. A mouse genomic clone of approximately 5 kb was obtained to determine the molecular basis for the regulation of the expression of mouse C-reactive protein. Several cis-acting elements in the 5′ flanking region that potentially regulate transcription were identified: two glucocorticoid-responsive elements, two CCAAT-enhancer-binding protein C (C/EBP) consensus elements that are required for the interleukin-1 responsiveness of some acute-phase reactant genes, an interleukin-6-responsive element, two hepatocyte nuclear factor-1 (HNF-1) elements and a single heat-shock element. Transfection of the hepatoma cell line Hep 3B.2 with a pCAT expression vector containing the 5′ flanking sequence from -1083 to -3 bp from the transcriptional start site, and truncations of this sequence, localized elements that control the tissue-specific expression of mouse C-reactive protein to the two HNF-1 elements and a C/EBP, interleukin-1-responsive element located between -220 and -153, and -90 and -50 bp from the transcriptional start site. A constitutive nuclear protein from mouse-liver hepatocytes specifically binds to the HNF-1 elements. These findings explain the tissue-specific expression of the gene, as well as its limited expression during the acute-phase response.

Factors involved in control of tissue-specific expression of albumin gene

Cell ◽  
1987 ◽  
Vol 50 (4) ◽  
pp. 627-638 ◽  
Author(s):  
Silvia Cereghini ◽  
Michel Raymondjean ◽  
Alejandro Garcia Carranca ◽  
Philippe Herbomel ◽  
Moshe Yaniv
Keyword(s):  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Albumin Gene

scholarly journals Transcriptional regulation of the rat platelet factor 4 gene: interaction between an enhancer/silencer domain and the GATA site.

1991 ◽  
Vol 11 (12) ◽  
pp. 6116-6127 ◽  
Author(s):  
K Ravid ◽  
T Doi ◽  
D L Beeler ◽  
D J Kuter ◽  
R D Rosenberg
Keyword(s):  
Tissue Specificity ◽  
Bone Marrow Cells ◽  
Core Promoter ◽  
Transcriptional Start Site ◽  
Human Growth ◽  
Platelet Factor 4 ◽  
Upstream Region ◽  
Growth Hormone Gene ◽  
Specific Expression ◽  
Platelet Factor

We used various segments of the 5' upstream region of the rat platelet factor 4 (PF4) gene coupled to the human growth hormone gene and heterologous promoters to identify domains which are critical for tissue-specific expression. Transient expression experiments with rat bone marrow cells and other cell lines revealed a complex interplay between a core promoter domain from -97 to the transcriptional start site and an enhancer/silencer domain from -448 to -112. The core promoter contains a GATA site at -31 to -28 whose mutation to TATA or AATA decreases tissue specificity and moderately affects expression in megakaryocytes as well as a positively acting subdomain from -97 to -83 whose removal decreases overall transcription without affecting tissue specificity. The enhancer/silencer domain possesses three positively acting subdomains from -380 to -362, -270 to -257, and -137 to -120 as well as a negatively acting subdomain at -184 to -151 which is able to reduce overall transcription but has no effect on tissue specificity. The subdomain from -380 to -362 is most critical in restricting gene expression driven either by the PF4 promoter or by a heterologous promoter to the megakaryocytic lineage. The subdomains from -270 to -257 and -137 to -120 function together with the subdomain from -380 to -362 to somewhat increase tissue specificity. Simultaneous mutation of the GATA site and deletion of either the whole enhancer/silencer domain or the subdomain from -380 to -362 or -137 to -120 reduce transcription in megakaryocytes by 10- to 30-fold. On the basis of the above-described results, we propose that the megakaryocyte-specific enhancer/silencer domain and the GATA site are responsible for high-level expression of the PF4 gene in a lineage-specific manner.

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