Genetic dissection of dome formation in a mammary cell line: identification of two genes with opposing action.

In this work, we extend the study of the genes controlling the formation of domes in the rat mammary cell line LA7 under the influence of DMSO. The role of the rat8 gene has already been demonstrated. We have now studied two additional genes. The first, called 133, is the rat ortholog of the human epithelial membrane protein 3 (EMP3), a member of the peripheral myelin protein 22 (PMP22)/EMP/lens-specific membrane protein 20 (MP20) gene family that encodes for tetratransmembrane proteins; it is expressed in the LA7 line in the absence of DMSO but not in its presence. The second gene is the beta subunit of the amiloride-sensitive Na(+) channel. Studies with antisense oligonucleotides show that the formation of domes is under the control of all three genes: the expression of rat8 is required for both their formation and their persistence; the expression of the Na(+) channel beta subunit is required for their formation; and the expression of gene 133 blocks the expression of the Na(+) channel genes, thus preventing formation of the domes. The formation of these structures is also accompanied by the expression of alpha(6)beta(1) integrin, followed by that of E-cadherin and cytokeratin 8. It appears, therefore, that dome formation requires the activity of the Na(+) channel and the rat8-encoded protein and is under the negative control of gene 133. DMSO induces dome formation by blocking this control.