The aim of the present study was to identify the functional domains of the upstream region of the rabbit whey acidic protein (WAP) gene, which has been used with considerable efficacy to target the expression of several foreign genes to the mammary gland. We have shown that this region exhibits three sites hypersensitive to DNase I digestion in the lactating mammary gland, and that all three sites harbour elements which can bind to Stat5 in vitro in bandshift assays. However, not all hypersensitive regions are detected at all stages from pregnancy to weaning, and the level of activated Stat5 detected in the rabbit mammary gland is low except during lactation. We have studied the role of the distal site, which is only detected during lactation, in further detail. It is located within a 849bp region that is required to induce a strong expression of the chloramphenicol acetyltransferase reporter gene in transfected mammary cells. Taken together, these results suggest that this region, centred around a Stat5-binding site and surrounded by a variable chromatin structure during the pregnancy–lactation cycle, may play a key role in regulating the expression of this gene in vivo. Furthermore, this distal region exhibits sequence similarity with a region located around 3kb upstream of the mouse WAP gene. The existence of such a distal region in the mouse WAP gene may explain the differences in expression between 4.1 and 2.1kb mouse WAP constructs.
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November 2001
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Research Article|
October 25 2001
A distal region, hypersensitive to DNase I, plays a key role in regulating rabbit whey acidic protein gene expression
Benjamin MILLOT;
Benjamin MILLOT
∗Unité de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, 78 352 Jouy en josas Cedex, France
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Marie-Louise FONTAINE;
Marie-Louise FONTAINE
∗Unité de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, 78 352 Jouy en josas Cedex, France
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Dominique THEPOT;
Dominique THEPOT
†Unité de Biologie du Développement et Biotechnologies, Institut National de la Recherche Agronomique, 78 352 Jouy en josas Cedex, France
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Eve DEVINOY
Eve DEVINOY
1
∗Unité de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, 78 352 Jouy en josas Cedex, France
1To whom correspondence should be addressed (e-mail devinoy@jouy.inra.fr).
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Publisher: Portland Press Ltd
Received:
July 23 2001
Accepted:
August 16 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 359 (3): 557–565.
Article history
Received:
July 23 2001
Accepted:
August 16 2001
Citation
Benjamin MILLOT, Marie-Louise FONTAINE, Dominique THEPOT, Eve DEVINOY; A distal region, hypersensitive to DNase I, plays a key role in regulating rabbit whey acidic protein gene expression. Biochem J 1 November 2001; 359 (3): 557–565. doi: https://doi.org/10.1042/bj3590557
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