Overexpression of the Na+,K+-ATPase α1 subunit increases Na+,K+-ATPase function in A549 cells

Phillip Factor*, Christina Senne, Vidas Dumasius, Karen Ridge, H. Ari Jaffe, Bruce Uhal, Zahn Gao, J. Iasha Sznajder

*Corresponding author for this work

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

We hypothesized that viral mediated transfer of Na+,K+-ATPase subunit genes to alveolar epithelial cells to overexpress Na+,K+-ATPase could increase Na+,K+-ATPase function. We produced replication-deficient human type 5 adenoviruses that contained cytomegalovirus (CMV)-driven cDNAs for the rat α1, and β1 subunits of Na+,K+-ATPase (AdMRCMVα1 and AdMRCMVβ1, respectively). These viruses were used to transduce human adenocarcinoma cells (A549) in culture. Na+,K+-ATPase function was increased by 2.5-fold in the AdMRCMVα1-infected cells. Sham and AdMRCMVβ1-infected cells, and cells infected by a CMV-driven β-galactosidase-expressing adenovirus, had no increases in Na+,K+-ATPase activity. A549 cells infected with multiplicities of infection of 10-200 of AdMRCMVα1 demonstrated expression of a rat α1 mRNA and increased α1 protein; no change in β1 message or protein was noted. Ouabain sensitivity was measured in A549 cells following infection with AdMRCMVα1. In contrast to controls, AdMRCMVα1-infected cells demonstrated two IC50s. The first was similar to the IC50s of the controls: the second IC50 was 2 logs greater than the first, consistent with the presence of both the rat and human α1 isozymes. These results demonstrate for the first time that adenoviruses can be used to augment Na+,K+-ATPase function.

Original languageEnglish (US)
Pages (from-to)741-749
Number of pages9
JournalAmerican journal of respiratory cell and molecular biology
Volume18
Issue number6
DOIs
StatePublished - 1998

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

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