Home LiteratureArticle Details
PMID: 11672429 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A distal region, hypersensitive to DNase I, plays a key role in regulating rabbit whey acidic protein gene expression.

The Biochemical journal ·Vol. 359 ·No. Pt 3 ·2001-11-01 ·Pages 557-65

Millot B, Fontaine ML, Thepot D, Devinoy E

Abstract

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 849 bp 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 3 kb 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 2.1 kb mouse WAP constructs.

MeSH Terms
Animals Cells, Cultured DNA-Binding Proteins/metabolism Deoxyribonuclease I/metabolism Female Gene Expression Regulation Genes, Reporter Lactation/physiology Mammary Glands, Animal/physiology Milk Proteins/genetics,metabolism Pregnancy Protein Isoforms Rabbits Recombinant Fusion Proteins/metabolism Regulatory Sequences, Nucleic Acid/genetics STAT5 Transcription Factor Trans-Activators/metabolism Weaning
Chemicals
DNA-Binding Proteins Milk Proteins Protein Isoforms Recombinant Fusion Proteins STAT5 Transcription Factor Trans-Activators whey acidic proteins Deoxyribonuclease I
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Millot B
Unité de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, 78 352 Jouy en josas Cedex, France.
Fontaine M L
Thepot D
Devinoy E
References (31)
31 references, click to expand
  1. Transcriptional induction of stromelysin-3 in mesodermal cells is mediated by an upstream CCAAT/enhancer-binding protein element associated with a DNase I-hypersensitive site.
    J Biol Chem. 1999 Dec 24;274(52):37177-85 PMID: 10601280
  2. Expression of human erythropoietin transgenes and of the endogenous WAP gene in the mammary gland of transgenic rabbits during gestation and lactation.
    Transgenic Res. 1998 Jul;7(4):311-7 PMID: 9859219
  3. Differential regulation of gene activity and chromatin structure within the human serpin gene cluster at 14q32.1 in macrophage microcell hybrids.
    Nucleic Acids Res. 2000 Apr 15;28(8):1767-77 PMID: 10734196
  4. Assignment of the rabbit whey acidic protein gene (WAP) to rabbit chromosome 10 by in situ hybridization and description of a large region surrounding this gene.
    Cytogenet Cell Genet. 2000;89(1-2):107-9 PMID: 10894949
  5. Endogenous protein kinase CK2 participates in Wnt signaling in mammary epithelial cells.
    J Biol Chem. 2000 Aug 4;275(31):23790-7 PMID: 10806215
  6. The gene for a novel member of the whey acidic protein family encodes three four-disulfide core domains and is asynchronously expressed during lactation.
    J Biol Chem. 2000 Jul 28;275(30):23074-81 PMID: 10801834
  7. Lactogenic activity in the serum of rabbits during pregnancy and early lactation.
    J Endocrinol. 1977 Oct;75(1):33-42 PMID: 925587
  8. Role of prolactin the the expression of casein genes in the virgin rabbit.
    Cell Differ. 1979 Feb;8(1):49-59 PMID: 455473
  9. Mouse whey acidic protein is a novel member of the family of 'four-disulfide core' proteins.
    Nucleic Acids Res. 1982 Apr 24;10(8):2677-84 PMID: 6896234
  10. A camel milk whey protein rich in half-cystine. Primary structure, assessment of variations, internal repeat patterns, and relationships with neurophysin and other active polypeptides.
    Eur J Biochem. 1986 Aug 15;159(1):195-201 PMID: 3743571
  11. Lactogenic hormone and cell type-specific control of the whey acidic protein gene promoter in transfected mouse cells.
    Mol Endocrinol. 1991 Nov;5(11):1624-32 PMID: 1685765
  12. Rabbit whey acidic protein concentration in milk, serum, mammary gland extract, and culture medium.
    J Dairy Sci. 1991 Dec;74(12):4143-50 PMID: 1787186
  13. Hormone responsive elements within the upstream sequences of the rabbit whey acidic protein (WAP) gene direct chloramphenicol acetyl transferase (CAT) reporter gene expression in transfected rabbit mammary cells.
    Mol Cell Endocrinol. 1991 Oct;81(1-3):185-93 PMID: 1797585
  14. Developmental and environmental regulation of a mammary gland-specific nuclear factor essential for transcription of the gene encoding beta-casein.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3130-4 PMID: 1557422
  15. Glucocorticoid receptor binding sites in the promoter region of milk protein genes.
    J Steroid Biochem Mol Biol. 1993 Dec;47(1-6):75-81 PMID: 8274444
  16. Variation of transferrin mRNA concentration in the rabbit mammary gland during the pregnancy-lactation-weaning cycle and in cultured mammary cells. A comparison with the other major milk protein mRNAs.
    Eur J Endocrinol. 1994 May;130(5):522-9 PMID: 8180682
  17. Participation of Ets transcription factors in the glucocorticoid response of the rat tyrosine aminotransferase gene.
    Mol Cell Biol. 1994 Jun;14(6):4116-25 PMID: 7910945
  18. Involvement of Ets-related proteins in hormone-independent mammary cell-specific gene expression.
    Eur J Biochem. 1994 Aug 1;223(3):997-1006 PMID: 8055977
  19. Regulation of human protein C gene expression by the mouse WAP promoter.
    Transgenic Res. 1994 Nov;3(6):335-43 PMID: 8000430
  20. Glucocorticoid regulation of rat whey acidic protein gene expression involves hormone-induced alterations of chromatin structure in the distal promoter region.
    Mol Endocrinol. 1994 Oct;8(10):1328-35 PMID: 7854350
  21. Regulation of the sheep beta-lactoglobulin gene by lactogenic hormones is mediated by a transcription factor that binds an interferon-gamma activation site-related element.
    Mol Endocrinol. 1994 Nov;8(11):1528-36 PMID: 7877621
  22. Nuclear factor I and mammary gland factor (STAT5) play a critical role in regulating rat whey acidic protein gene expression in transgenic mice.
    Mol Cell Biol. 1995 Apr;15(4):2063-70 PMID: 7891701
  23. A combination of distal and proximal regions is required for efficient prolactin regulation of transfected rabbit alpha s1-casein chloramphenicol acetyltransferase constructs.
    Mol Endocrinol. 1994 Dec;8(12):1720-30 PMID: 7677833
  24. Regulation of ovine beta-lactoglobulin gene expression during the first stage of lactogenesis.
    Biochem Biophys Res Commun. 1995 Apr 26;209(3):1089-93 PMID: 7733963
  25. Position-independent expression of whey acidic protein transgenes.
    J Biol Chem. 1995 May 12;270(19):11119-29 PMID: 7744742
  26. Production of human surfactant protein C in milk of transgenic mice.
    Transgenic Res. 1995 Jul;4(4):232-40 PMID: 7655513
  27. Regulation of mammary gland factor/Stat5a during mammary gland development.
    Mol Endocrinol. 1995 Nov;9(11):1598-609 PMID: 8584036
  28. Rabbit whey acidic protein gene upstream region controls high-level expression of bovine growth hormone in the mammary gland of transgenic mice.
    Mol Reprod Dev. 1995 Nov;42(3):261-7 PMID: 8579839
  29. An Ets site in the whey acidic protein gene promoter mediates transcriptional activation in the mammary gland of pregnant mice but is dispensable during lactation.
    Mol Endocrinol. 1995 Jun;9(6):717-24 PMID: 8592517
  30. Temporal profiles of appearance of DNase I hypersensitive sites associated with the ovine beta-lactoglobulin gene differ in sheep and transgenic mice.
    Mol Gen Genet. 1998 Apr;257(6):649-54 PMID: 9604888
  31. High expression of the human hepatocarcinoma-intestine-pancreas/pancreatic-associated protein (HIP/PAP) gene in the mammary gland of lactating transgenic mice. Secretion into the milk and purification of the HIP/PAP lectin.
    Eur J Biochem. 2000 Mar;267(6):1665-71 PMID: 10712597
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/0264-6021:3590557
Published
2001-11-01
Pages
557-65
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1222176
Subset
IM
External Links
PubMed source DOI: 10.1042/0264-6021:3590557 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2