Proteomic dissection of dome formation in a mammary cell line: Role of tropomyosin-5b and maspin
- 8 May 2001
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 98 (10) , 5608-5613
- https://doi.org/10.1073/pnas.091101898
Abstract
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 beta-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 beta-subunit expression in the absence of DMSO. Therefore, dome formation requires the expression of tropomyosin-5b, in addition to the ENaC beta-subunit and the rat8 proteins, and is under the negative control of maspin.Keywords
This publication has 50 references indexed in Scilit:
- Melanie II – a third‐generation software package for analysis of two‐dimensional electrophoresis images: II. AlgorithmsElectrophoresis, 1997
- Epithelial membrane protein-2 and epithelial membrane protein-3: two novel members of the peripheral myelin protein 22 gene familyGene, 1996
- Restoration of microfilament bundle organization in v-raf-transformed NRK cells after transduction with tropomyosin 2 cDNACancer Letters, 1994
- The focusing positions of polypeptides in immobilized pH gradients can be predicted from their amino acid sequencesElectrophoresis, 1993
- Coordinated expression of extracellular matrix-degrading proteinases and their inhibitors regulates mammary epithelial function during involution.The Journal of cell biology, 1992
- Comparative two-dimensional gel analysis and microsequencing identifies gelsolin as one of the most prominent downregulated markers of transformed human fibroblast and epithelial cells.The Journal of cell biology, 1990
- Two‐dimensional electrophoresis. The current state of two‐dimensional electrophoresis with immobilized pH gradientsElectrophoresis, 1988
- Differentiation and morphogenesis of mammary cellsin vitroProceedings of the Royal Society of London. B. Biological Sciences, 1980
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976