Interactions between the rabbit CSN1 gene and the nuclear matrix of stably transfected HC11 mammary epithelial cells vary with its level of expression.
The expression of casein genes is specific to the mammary gland and maximal during lactation. However, among the numerous mammary cell lines described so far, only a few express some casein genes. The regulatory regions of casein genes have been largely described but the mechanisms explaining the mammary specific expression of these genes, and their silencing in most mammary cell lines, have not yet been fully elucidated. To test the hypothesis that the nuclear location of the casein genes may affect their expression, we transfected HC11 mouse mammary cell line with a 100 kb DNA fragment surrounding the rabbit alpha S1 casein gene. We derived stable clones which express or not the transfected rabbit casein gene, in the same cellular context, independently of the number of transgene copies. Metaphase spreads were prepared from the different clones and the transfected genes were localized. Unexpectedly, we observed that in the original HC11 cell line the number of chromosomes per metaphase spread is close to 80, suggesting that HC11 cells have undergone a duplication event, since the mouse karyotype is 2n = 40. In alpha S1 casein expressing cells, the expression level does not clearly correlate with a localization of the transfected DNA proximal to the centromeres or the telomeres. Analysis of the localization of the transfected DNA in nuclear halos allows us to conclude that when expressed, transfected DNA is more closely linked to the nuclear matrix. The next step will be to study the attachment of the endogenous casein gene in mammary nuclei during lactation.
Poussin
Karine
K
UMR 8646 CNRS, U565 INSERM, Département Régulations, Développement et Diversité Moléculaire, Muséum National d'Histoire Naturelle, Case Postale 26, 57 rue Cuvier, 75231 Paris cedex 05, France.
Hayes
Hélène
H
Pauloin
Alain
A
Chanat
Eric
E
Fontaine
Marie-Louise
ML
Aujean
Etienne
E
Sun
Jian-Sheng
JS
Debey
Pascale
P
Devinoy
Eve
E
D000818:Animals; D002364:Caseins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002875:Chromosomes; D004247:DNA / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D018628:Gene Dosage; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D008321:Mammary Glands, Animal / Q000166:cytology*; D051379:Mice; D015530:Nuclear Matrix / Q000378:metabolism*; D011817:Rabbits; D019076:Transgenes
D002364:Caseins; D004247:DNA