Proteomic Dissection of Dome Formation in a Mammary Cell Line: Role of Tropomyosin-5b and Maspin

• Published in: Proceedings of the National Academy of Sciences - PNAS 2001-05, Vol.98 (10), p.5608-5613
• Main Authors: Zucchi, I., Bini, L., Valaperta, R., Ginestra, A., Albani, D., Susani, L., Sanchez, J. C., Liberatori, S., Magi, B., Raggiaschi, R., Hochstrasser, D. F., Pallini, V., Vezzoni, P., Dulbecco, R.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antisense oligonucleotides; Base Sequence; Biological Sciences; Blotting, Northern; Cell lines; Cells; Complementary DNA; Cultured cells; DNA Primers; Electrophoresis; Epithelial Sodium Channels; Gels; Genes; Genes, Tumor Suppressor; Genetics; Mammary glands; Mammary Glands, Animal - cytology; Mammary Glands, Animal - metabolism; Messenger RNA; Proteins - antagonists & inhibitors; Proteins - genetics; Proteins - physiology; Proteome; Rats; Rats, Sprague-Dawley; RNA, Messenger - genetics; Serpins - genetics; Serpins - physiology; Sodium Channels - metabolism; Structural domes; Tropomyosin - genetics; Tropomyosin - physiology; Tumor Cells, Cultured
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.091101898

In this work we extended the study of genes controlling the formation of specific differentiation structures called "domes" formed by the rat mammary adenocarcinoma cell line LA7 under the influence of DMSO. We have reported previously that an interferon-inducible gene, rat-8, and the β-subunit of the epithelial sodium channel (ENaC) play a fundamental role in this process. Now, we used a proteomic approach to identify proteins differentially expressed either in DMSO-induced LA7 or in 106A10 cells. Two differentially expressed proteins were investigated. The first, tropomyosin-5b, strongly expressed in DMSO-induced LA7 cells, is needed for dome formation because its synthesis inhibition by the antisense RNA technology abolished domes. The second protein, maspin, strongly expressed in the uninduced 106A10 cell line, inhibits dome formation because 106A10 cells, transfected with rat8 cDNA (the function of which is required for the organization of these structures), acquired the ability to develop domes when cultured in presence of an antimaspin antibody. Dome formation in these cultures are accompanied by ENaC β-subunit expression in the absence of DMSO. Therefore, dome formation requires the expression of tropomyosin-5b, in addition to the ENaC β-subunit and the rat8 proteins, and is under the negative control of maspin.