Nuclear protein factors binding to a class I patatin promoter region are tuber-specific and sucrose-inducible

1994 ◽  
Vol 26 (2) ◽  
pp. 603-615 ◽  
Author(s):  
Soo Young Kim ◽  
Gregory D. May ◽  
William D. Park
Keyword(s):  
Promoter Region ◽  
Nuclear Protein ◽  
Class I ◽  
Patatin Promoter ◽  
Class I Patatin

Identification of a cell-type-specific transcriptional repressor in the promoter region of the mouse hepatocyte growth factor gene

1994 ◽  
Vol 14 (11) ◽  
pp. 7046-7058
Author(s):  
Y Liu ◽  
A B Beedle ◽  
L Lin ◽  
A W Bell ◽  
R Zarnegar
Keyword(s):  
Epithelial Cells ◽  
Promoter Region ◽  
Nuclear Protein ◽  
Transcriptional Repressor ◽  
Cell Type ◽  
Mobility Shift ◽  
A Cell ◽  
Cell Type Specific ◽  
Gel Mobility Shift ◽  
Hepatocyte Growth

Hepatocyte growth factor (HGF), a cytokine with multiple functions, exhibits cell-type-specific as well as cytokine- and steroid hormone-regulated expression. The HGF gene is known to be expressed predominately in mesenchymal but not in epithelial cells. In this study, we report the identification of a cell-type-specific transcriptional repressor in the promoter region of the mouse HGF gene, which is evidently responsible for the suppression of HGF expression in epithelial cells. Gel mobility shift assays and DNase I footprinting studies revealed that a 27-bp element (-16 to +11) around the transcription initiation site is responsible for the binding of a nuclear protein which is present in epithelial but not in mesenchymally derived cells. Further analysis of the binding activity of the DNA region with nuclear protein revealed that an approximately 19-bp sequence containing a unique palindromic structure (5'-AACCGACCGGTT-3') overlapped by a CAP box is essential for binding. Substitution of a single base (the contact site) within this region by site-directed mutagenesis resulted in total abrogation of the binding of the nuclear protein and a concomitant increase in the transcriptional activity of various lengths of HGF-chloramphenicol acetyltransferase fused genes when transfected into the epithelial cell line RL95-2 but not the mesenchymal cell line NIH 3T3. Southwestern (DNA-protein) analyses revealed that the nuclear protein which binds to this repressor element is a single polypeptide of approximately 70 kDa. Analysis of the nuclear extract prepared from regenerating mouse liver at various times after two-thirds partial hepatectomy by gel mobility shift assay revealed a substantial reduction (more than 75% within 3 h) in the binding of the repressor to its cognate binding site. Our results suggest that a cis-acting transcriptional repressor in the promoter region of the mouse HGF gene is involved in cell-type-specific regulation through binding to its cognate trans-acting protein which exists in epithelial cells but is absent in fibroblast cells.

scholarly journals Identification of a cell-type-specific transcriptional repressor in the promoter region of the mouse hepatocyte growth factor gene.

1994 ◽  
Vol 14 (11) ◽  
pp. 7046-7058 ◽  
Author(s):  
Y Liu ◽  
A B Beedle ◽  
L Lin ◽  
A W Bell ◽  
R Zarnegar
Keyword(s):  
Epithelial Cells ◽  
Promoter Region ◽  
Nuclear Protein ◽  
Transcriptional Repressor ◽  
Cell Type ◽  
Mobility Shift ◽  
A Cell ◽  
Cell Type Specific ◽  
Gel Mobility Shift ◽  
Hepatocyte Growth

Hepatocyte growth factor (HGF), a cytokine with multiple functions, exhibits cell-type-specific as well as cytokine- and steroid hormone-regulated expression. The HGF gene is known to be expressed predominately in mesenchymal but not in epithelial cells. In this study, we report the identification of a cell-type-specific transcriptional repressor in the promoter region of the mouse HGF gene, which is evidently responsible for the suppression of HGF expression in epithelial cells. Gel mobility shift assays and DNase I footprinting studies revealed that a 27-bp element (-16 to +11) around the transcription initiation site is responsible for the binding of a nuclear protein which is present in epithelial but not in mesenchymally derived cells. Further analysis of the binding activity of the DNA region with nuclear protein revealed that an approximately 19-bp sequence containing a unique palindromic structure (5'-AACCGACCGGTT-3') overlapped by a CAP box is essential for binding. Substitution of a single base (the contact site) within this region by site-directed mutagenesis resulted in total abrogation of the binding of the nuclear protein and a concomitant increase in the transcriptional activity of various lengths of HGF-chloramphenicol acetyltransferase fused genes when transfected into the epithelial cell line RL95-2 but not the mesenchymal cell line NIH 3T3. Southwestern (DNA-protein) analyses revealed that the nuclear protein which binds to this repressor element is a single polypeptide of approximately 70 kDa. Analysis of the nuclear extract prepared from regenerating mouse liver at various times after two-thirds partial hepatectomy by gel mobility shift assay revealed a substantial reduction (more than 75% within 3 h) in the binding of the repressor to its cognate binding site. Our results suggest that a cis-acting transcriptional repressor in the promoter region of the mouse HGF gene is involved in cell-type-specific regulation through binding to its cognate trans-acting protein which exists in epithelial cells but is absent in fibroblast cells.

scholarly journals Sequence-specific interactions of a nuclear protein factor with the promoter region of a rice gene for α-amylase,RAmy3D

1994 ◽  
Vol 22 (11) ◽  
pp. 1948-1953 ◽  
Author(s):  
Shin-ichiro Mitsunaga ◽  
Raymond L. Rodriguez ◽  
Junji Yamaguchi
Keyword(s):  
Promoter Region ◽  
Nuclear Protein ◽  
Specific Interactions ◽  
Rice Gene ◽  
Protein Factor

The expression of class I patatin gene fusions in transgenic potato varies with both gene and cultivar

1991 ◽  
Vol 16 (1) ◽  
pp. 153-160 ◽  
Author(s):  
K. S. Blundy ◽  
M. A. C. Blundy ◽  
D. Carter ◽  
F. Wilson ◽  
W. D. Park ◽  
...  
Keyword(s):  
Transgenic Potato ◽  
Class I ◽  
Gene Fusions ◽  
Class I Patatin ◽  
Patatin Gene

Two Common Functional Polymorphisms in the Promoter Region of the Coagulation Factor VII Gene Determining Plasma Factor VII Activity and Mass Concentration

Blood ◽  
1999 ◽  
Vol 93 (10) ◽  
pp. 3432-3441 ◽  
Author(s):  
Ferdinand M. van ’t Hooft ◽  
Angela Silveira ◽  
Per Tornvall ◽  
Anastasia Iliadou ◽  
Ewa Ehrenborg ◽  
...  
Keyword(s):  
Promoter Region ◽  
Nuclear Protein ◽  
Plasma Concentrations ◽  
Coagulation Factor ◽  
Protein S ◽  
Factor Vii ◽  
Binding Properties ◽  
Coagulation Factor Vii ◽  
Functional Polymorphisms ◽  
Plasma Factor

Recent studies have provided evidence for associations between common polymorphic markers in the coagulation factor VII (FVII) gene and plasma FVII levels. Here we describe two common, nonrelated, functional polymorphisms in the promoter region of the FVII gene, a G to T substitution at position −401 and a novel G to A substitution at position −402. Both polymorphisms strongly influence the binding properties of nuclear protein(s). The rare −401T allele is associated with a reduced basal rate of transcription of the FVII gene in human hepatoblastoma cells and with reduced plasma concentrations of total FVII (VIIag) and fully activated FVII molecules (VIIa). In contrast, the rare −402A allele confers increased transcriptional activity and is associated with increased plasma FVII levels. Together, the two polymorphisms explained 18% and 28% of the variation in VIIag and VIIa, respectively, in a group of 183 healthy, middle-aged men. It is concluded that these polymorphisms are important for the regulation of the plasma levels of FVII and that they are likely to be useful genetic markers to resolve the issue of whether a causal relationship exists between FVII levels and risk of coronary heart disease.

scholarly journals Partial purification of a nuclear protein that binds to the CCAAT box of the mouse alpha 1-globin gene.

1986 ◽  
Vol 6 (3) ◽  
pp. 821-832 ◽  
Author(s):  
R B Cohen ◽  
M Sheffery ◽  
C G Kim
Keyword(s):  
Promoter Region ◽  
Nuclear Protein ◽  
Specific Binding ◽  
Globin Gene ◽  
Dimethyl Sulfate ◽  
Erythroid Cell ◽  
Partial Purification ◽  
Binding Studies ◽  
Murine Erythroleukemia ◽  
Ccaat Box

We enriched a fraction from nuclear extracts of murine erythroleukemia cells which contains a protein able to form stable complexes with the promoter region of the alpha 1-globin gene. Binding activity, which is present in mouse brain and a variety of cultured mouse and human cell lines, is not erythroid cell specific. Binding studies with alpha-globin gene promoter deletion mutants as well as DNase I footprinting and dimethyl sulfate protection studies showed that the factor bound specifically to the CCAAT box of the alpha 1 promoter. Enriched factor preparations exhibited weak binding to the promoter region of the beta maj-globin gene. This suggests that this protein could bind differentially to these two promoters in vivo. The enriched factor may be a ubiquitous nuclear protein involved in the differential regulation of the alpha 1- and beta maj-globin genes.

Functional analysis of a class I patatin gene SK24-1 in microtuber formation of transgenic potatoes

2008 ◽  
Vol 88 (4) ◽  
pp. 593-598 ◽  
Author(s):  
Huaijun Si ◽  
Jun Liu ◽  
Jian Huang ◽  
Conghua Xie
Keyword(s):  
Escherichia Coli ◽  
Cdna Clone ◽  
Antisense Rna ◽  
Cauliflower Mosaic Virus ◽  
Class I ◽  
35S Promoter ◽  
Transformed Plants ◽  
Transgenic Potatoes ◽  
Class I Patatin

Expression of a class I patatin cDNA clone, SK24-1, in Escherichia coli revealed that the cDNA clone possessed lipid acyl hydrolase (LAH) activity. Transformed potato plants were obtained via Agrobacterium-mediated transformation using the chimeric constructs containing the sense and antisense cDNA under the control cauliflower mosaic virus 35S (CaMV 35S) promoter. In some sense transformed plants, both sense patatin RNA and LAH activity were increased and further resulted in a significant increase of percentage of plantlets that formed microtubers and numbers of microtubers per plantlet in vitro. All antisense plants displayed a reduction in LAH activity. Both sense and antisense RNA could be detected in antisense plants, but transcripts of antisense RNA resulted in a reduction of endogenous sense RNA. Moreover, expression of antisense cDNA in some antisense transformed plants led to a significant decrease in the number of microtubers formed. These results suggest that SK24-1 was involved in regulating microtuber formation. Key words: Patatin, potato, Escherichia coli, sense RNA, antisense RNA

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