scholarly journals Interaction of a tissue-specific factor with an essential rat growth hormone gene promoter element.

1987 ◽  
Vol 7 (3) ◽  
pp. 1193-1197 ◽  
Author(s):  
B L West ◽  
D F Catanzaro ◽  
S H Mellon ◽  
P A Cattini ◽  
J D Baxter ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Dna Sequences ◽  
Site Directed Mutagenesis ◽  
Specific Factor ◽  
Growth Hormone Gene ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Rat Growth

Rat growth hormone (rGH) gene expression is normally restricted to the anterior pituitary. As a model of this tissue specificity, we compared the transient expression of an rGH-chloramphenicol acetyltransferase (CAT) hybrid gene in rGH-producing rat pituitary tumor (GC) cells and in non-rGH-producing rat fibroblast (rat-2) cells. Deletion analysis of the rGH portion of this hybrid gene demonstrated that DNA sequences within 140 base pairs 5' to the rGH gene were sufficient for correct cell type-specific expression. Deletion of an additional 35 base pairs of the rGH 5'-flanking DNA resulted in a loss of expression of the transfected hybrid gene and correlated with the interaction of a putative trans-acting factor with this region of the rGH promoter. This factor was detectable by DNase I footprinting in a crude nuclear extract from GC cells but not from rat-2 cells. Site-directed mutagenesis of the footprint region caused complete loss of expression of a hybrid gene containing 530 base pairs 5' to the rGH gene. Thus, the interaction of this factor, which we term GC2, is likely to be essential for the tissue-specific expression of the rGH gene.

Interaction of a tissue-specific factor with an essential rat growth hormone gene promoter element

1987 ◽  
Vol 7 (3) ◽  
pp. 1193-1197
Author(s):  
B L West ◽  
D F Catanzaro ◽  
S H Mellon ◽  
P A Cattini ◽  
J D Baxter ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Dna Sequences ◽  
Site Directed Mutagenesis ◽  
Specific Factor ◽  
Growth Hormone Gene ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Rat Growth

Rat growth hormone (rGH) gene expression is normally restricted to the anterior pituitary. As a model of this tissue specificity, we compared the transient expression of an rGH-chloramphenicol acetyltransferase (CAT) hybrid gene in rGH-producing rat pituitary tumor (GC) cells and in non-rGH-producing rat fibroblast (rat-2) cells. Deletion analysis of the rGH portion of this hybrid gene demonstrated that DNA sequences within 140 base pairs 5' to the rGH gene were sufficient for correct cell type-specific expression. Deletion of an additional 35 base pairs of the rGH 5'-flanking DNA resulted in a loss of expression of the transfected hybrid gene and correlated with the interaction of a putative trans-acting factor with this region of the rGH promoter. This factor was detectable by DNase I footprinting in a crude nuclear extract from GC cells but not from rat-2 cells. Site-directed mutagenesis of the footprint region caused complete loss of expression of a hybrid gene containing 530 base pairs 5' to the rGH gene. Thus, the interaction of this factor, which we term GC2, is likely to be essential for the tissue-specific expression of the rGH gene.

Melanocyte-specific gene expression: role of repression and identification of a melanocyte-specific factor, MSF

1994 ◽  
Vol 14 (5) ◽  
pp. 3494-3503
Author(s):  
U Yavuzer ◽  
C R Goding
Keyword(s):  
Gene Expression ◽  
Mutational Analysis ◽  
Site Directed Mutagenesis ◽  
Specific Factor ◽  
Specific Gene ◽  
Binding Motif ◽  
Specific Expression ◽  
Tissue Specific ◽  
Specific Gene Expression

For a gene to be transcribed in a tissue-specific fashion, expression must be achieved in the appropriate cell type and also be prevented in other tissues. As an approach to understanding the regulation of tissue-specific gene expression, we have analyzed the requirements for melanocyte-specific expression of the tyrosinase-related protein 1 (TRP-1) promoter. Positive regulation of TRP-1 expression is mediated by both an octamer-binding motif and an 11-bp element, termed the M box, which is conserved between the TRP-1 and other melanocyte-specific promoters. We show here that, consistent with its ability to activate transcription in a non-tissue-specific fashion, the M box binds the basic-helix-loop-helix factor USF in vitro. With the use of a combination of site-directed mutagenesis and chimeric promoter constructs, additional elements involved in regulating TRP-1 expression were identified. These include the TATA region, which appears to contribute to the melanocyte specificity of the TRP-1 promoter. Mutational analysis also identified two repressor elements, one at the start site, the other located at -240, which function both in melanoma and nonmelanoma cells. In addition, a melanocyte-specific factor, MSF, binds to sites which overlap both repressor elements, with substitution mutations demonstrating that binding by MSF is not required for repression. Although a functional role for MSF has not been unequivocally determined, the location of its binding sites leads us to speculate that it may act as a melanocyte-specific antirepressor during transcription of the endogenous TRP-1 gene.

Growth Hormone Gene Expression: Hormonal Regulation and Tissue-Specific Expression

1988 ◽  
pp. 3-12
Author(s):  
Norman L. Eberhardt ◽  
Peter A. Cattini ◽  
Linda N. Peritz ◽  
John D. Baxter ◽  
Randy Isaacs ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Hormonal Regulation ◽  
Growth Hormone Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression

scholarly journals A Pituitary-Specific Factor Interacts with an Upstream Promoter Element in the Rat Growth Hormone Gene*

1987 ◽  
Vol 1 (1) ◽  
pp. 90-96 ◽  
Author(s):  
Daniel F. Catanzaro ◽  
Brian L. West ◽  
John D. Baxter ◽  
Tim L. Reudelhuber
Keyword(s):  
Growth Hormone ◽  
Specific Factor ◽  
Growth Hormone Gene ◽  
Promoter Element ◽  
Upstream Promoter ◽  
Rat Growth
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