scholarly journals Deletion analyses of 5'-flanking region of the human elastin gene. Delineation of functional promoter and regulatory cis-elements.

1990 ◽  
Vol 265 (16) ◽  
pp. 9485-9490
Author(s):  
V M Kähäri ◽  
M J Fazio ◽  
Y Q Chen ◽  
M M Bashir ◽  
J Rosenbloom ◽  
...  
1997 ◽  
Vol 11 (5) ◽  
pp. 638-649 ◽  
Author(s):  
Colin D. Clyne ◽  
Yin Zhang ◽  
Liliya Slutsker ◽  
J. Michael Mathis ◽  
Perrin C. White ◽  
...  

Abstract Aldosterone synthase is a mitochondrial enzyme that catalyzes the conversion of 11-deoxycorticosterone to the potent mineralocorticoid aldosterone. The gene encoding aldosterone synthase, CYP11B2, is expressed in the zona glomerulosa of the adrenal cortex. Although the major physiological regulators of aldosterone production are angiotensin II (ANG II) and potassium (K+), the mechanisms by which these compounds regulate CYP11B2 transcription are unknown. Therefore we analyzed the human CYP11B2 5′-flanking region using a transient transfection expression system in the H295R human adrenocortical cell line. ANG II and K+ increased expression of a luciferase reporter construct containing 2015 bp of human CYP11B2 5′-flanking DNA. This response was mimicked by treatment with the calcium channel activator BAYK8644, whereas activation of the protein kinase C pathway with 12-o-tetradecanoylphorbol-13-acetate had no effect. Reporter gene activity was also increased after activation of cAMP-dependent pathways by (Bu)2cAMP. Deletion, mutation, and deoxyribonuclease I footprinting analyses of the CYP11B2 5′-flanking region identified two distinct elements at positions −71/−64 (TGACGTGA) and −129/−114 (CTCCAGCCTTGACCTT) that were both required for full basal reporter gene activity and for maximal induction by either cAMP or calcium-signaling pathways. The −71/−64 element, which resembles a consensus cAMP response element (CRE), bound CRE-binding proteins from H295R cell nuclear extracts as determined by electrophoretic mobility shift analysis. Analysis of the −129/−114 element using electrophoretic mobility shift analysis demonstrated binding of the orphan nuclear receptors steroidogenic factor 1 and chicken ovalbumin upstream promoter transcription factor. These data demonstrate that ANG II, K+, and cAMP-signaling pathways utilize the same SF-1 and CRE-like cis-elements to regulate human CYP11B2 expression.


1990 ◽  
Vol 94 (2) ◽  
pp. 191-196 ◽  
Author(s):  
Michael J Fazio ◽  
Veli-Matti Kähäri ◽  
Muhammad M Bashir ◽  
Biagio Saitta ◽  
Joel Rosenbloom ◽  
...  

1993 ◽  
Vol 70 (03) ◽  
pp. 500-505 ◽  
Author(s):  
B Wyler ◽  
L Daviet ◽  
H Bortkiewicz ◽  
J-C Bordet ◽  
J L McGregor

SummaryGlycoprotein CD36, also known as GPIIIb or GPIV, is a major platelet glycoprotein that bears the newly identified Naka alloantigen. The aim of this study was to clone platelet CD36 and investigate other forms of CD36-cDNA present in monocytes, endothelial and HEL cells. RNA from above mentioned cells were reverse transcribed (RT), using specific primers for CD36, and amplified by the polymerase chain reaction (PCR) technique. Sequencing the different amplified platelet derived cDNA fragments, spanning the whole coding and flanking regions, showed the near identity between platelet and CD36-placenta cDNA. Platelet CD36-cDNA cross-hybridized, in Southern blots, with RT-PCR amplified cDNA originating from monocytes, endothelial and HEL cells. However, monocytes showed a RT-PCR amplified cDNA fragment (561 bp) that was present in platelets and placenta but not on endothelial on HEL-cells. Northern blot analysis of platelet RNA hybridized with placenta CD36 indicated the presence of a major (1.95 kb) and a minor (0.95 kb) transcript. The 1.95 kb transcript was the only one observed on Northern blots of monocytes, endothelial and HEL cells. These results indicate that the structure of CD36 expressed in platelets is similar, with the exception of the 3’ flanking region, to that of placenta. Differences in apparent molecular weight between CD36 and CD36-like glycoproteins may be due to post-translational modifications.


Diabetes ◽  
1985 ◽  
Vol 34 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S. Elbein ◽  
P. Rotwein ◽  
M. A. Permutt ◽  
G. I. Bell ◽  
N. Sanz ◽  
...  

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