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Volume 271, Number 38, Issue of September 20, 1996 pp. 23211-23221
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Translational Regulation of Na,K-ATPase alpha 1 and beta 1 Polypeptide Expression in Epithelial Cells

(Received for publication, March 12, 1996, and in revised form, June 6, 1996)

Kent K. Grindstaff , Gustavo Blanco and Robert W. Mercer

From the Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110

To investigate the regulation of the Na,K-ATPase, we have studied the expression of the Na,K-ATPase polypeptides in several mammalian cell lines using the vaccinia virus/T7 RNA polymerase expression system. Infection of several fibroblast-like cell lines with viral recombinants containing the Na,K-ATPase alpha  and beta  isoforms, the glucose transporters, GLUT 1 and GLUT 4, or the capsid protein of the Sindbis virus all result in the production of the appropriate protein products. However, all epithelial cell lines tested fail to synthesize the Na,K-ATPase viral recombinants, yet they efficiently express the other virally directed polypeptides. While Madin-Darby canine kidney (MDCK) epithelial cells infected with the Na,K-ATPase alpha 1 or beta 1 recombinant viruses produce both mRNAs, the messages are inefficiently translated. Furthermore, the RNA from infected MDCK cells does not direct the in vitro synthesis of the beta 1 polypeptide, whereas the message from infected fibroblast-like BSC 40 cells is efficiently translated both in vivo and in vitro. Moreover, the synthesis of the H,K-ATPase alpha  subunit is also limited in MDCK cells, although the H,K-ATPase beta  subunit is efficiently expressed. Expression of chimeras constructed between the Na+ pump beta 1 isoform and the H,K-ATPase beta  subunit indicates that sequences in the 5' coding region of the beta 1 message have an inhibitory effect; however, the stringent translational regulation of the beta 1 isoform in MDCK cells requires the 5' and 3' regions of the coding sequence. The ability of the polarized cell lines to limit the synthesis of the Na+ pump polypeptides while expressing other vaccinia recombinants at high levels suggests that the polarized cells possess a stringent mechanism for the specific translational regulation of a select set of messages.


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