Expression of an ouabain-resistant Na,K-ATPase in CV-1 cells after transfection with a cDNA encoding the rat Na,K-ATPase alpha 1 subunit.

We have used a gene transfer system to investigate the relationship between expression of the rat Na,K-ATPase alpha 1 subunit gene and ouabain-resistant Na,K-ATPase activity. A cDNA clone encoding the entire rat Na,K-ATPase alpha 1 subunit was inserted into the expression vector pSV2neo. This construct (pSV2 alpha 1) conferred resistance to 100 microM ouabain to ouabain-sensitive CV-1 cells. Hybridization analysis of transfected clones revealed the presence of both rat-specific and endogenous Na,K-ATPase alpha 1 subunit DNA and mRNA sequences. A single form of highly ouabain-sensitive 86Rb+ uptake was detected in CV-1 cells, whereas two distinct classes of ouabain-inhibitable uptake were observed in transfectants. One class exhibited the high ouabain sensitivity of the endogenous monkey Na,K-ATPase, while the second class showed the reduced ouabain sensitivity characteristic of the rodent renal Na,K-ATPase. Examination of the ouabain-sensitive, sodium-dependent ATPase activity of the transfectants also revealed a low affinity component of Na,K-ATPase activity characteristic of the rodent kidney enzyme. These results suggest that expression of the rat alpha 1 subunit gene is directly responsible for ouabain-resistant Na,K-ATPase activity in transfected CV-1 cells.

• Publication year

  1988

• Venue

  Journal of Biological Chemistry

• Publication date

  1988-06-05

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/s0021-9258(18)68559-xPMID 2836394
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• CV-1 cell recipients of the mouse ouabain resistance gene express a ouabain-insensitive Na,K-ATPase after growth in cardioactive steroids.

  1988cited by this paper
• Genes encoding alpha and beta subunits of Na,K-ATPase are located on three different chromosomes in the mouse.

  1987cited by this paper
• Ouabain resistance conferred by expression of the cDNA for a murine Na+, K+-ATPase alpha subunit.

  1987cited by this paper
• Multiple genes encode the human Na+,K+-ATPase catalytic subunit.

  1987cited by this paper
• Chromosome-mediated transfer of the murine Na,K-ATPase alpha subunit confers ouabain resistance

  1987cited by this paper
• Amplification of DNA sequences coding for the Na,K-ATPase alpha-subunit in ouabain-resistant C+ cells

  1986cited by this paper
• Use of monoclonal antibodies to culture rat proximal tubule cells.

  1986cited by this paper
• Molecular cloning and sequence analysis of human Na,K-ATPase beta-subunit.

  1986cited by this paper
• Molecular cloning of three distinct forms of the Na+,K+-ATPase alpha-subunit from rat brain.

  1986cited by this paper
• Molecular cloning of rat brain Na,K-ATPase alpha-subunit cDNA.

  1985cited by this paper
• Insulin affects the sodium affinity of the rat adipocyte (Na+,K+)-ATPase.

  1985cited by this paper
• Expression of an ouabain resistance gene in transfected cells. Ouabain treatment induces a K+-transport system.

  1985cited by this paper
• Enzymatic properties of separated isozymes of the Na,K-ATPase. Substrate affinities, kinetic cooperativity, and ion transport stoichiometry.

  1985cited by this paper
• Molecular cloning of the mouse ouabain-resistance gene.

  1984cited by this paper
• "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum.

  1984cited by this paper
• Assignment of the gene governing cellular ouabain resistance to Mus musculus chromosome 3 using human/mouse microcell hybrids

  1979cited by this paper
• Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.

  1979cited by this paper
• DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells.

  1979cited by this paper
• Two molecular forms of (Na+ + K+)-stimulated ATPase in brain. Separation, and difference in affinity for strophanthidin.

  1979cited by this paper
• (Na, K)ATPase activity in membrane preparations of ouabain-resistant HeLa cells.

  1977cited by this paper
• Ouabain as a selective agent in the isolation of somatic cell hybrids.

  1975cited by this paper
• Detection of specific sequences among DNA fragments separated by gel electrophoresis.

  1975cited by this paper
• Photoaffinity labeling of the ouabain-binding site on (Na+ plus K+) adenosinetriphosphatase.

  1974cited by this paper
• Isolation of ouabain-resistant human diploid fibroblasts.

  1974cited by this paper
• THE ACTION OF CARDIAC GLYCOSIDES ON ION MOVEMENTS.

  1964cited by this paper
• Protein measurement with the Folin phenol reagent.

  1951cited by this paper

• Uncovering key steps in FGF12 cellular release reveals a common mechanism for unconventional FGF protein secretion

  2024cites this paper
• Inhibition of Non Canonical HIV-1 Tat Secretion Through the Cellular Na+,K+-ATPase Blocks HIV-1 Infection

  2017cites this paper
• Cleistanthus collinus induces type I distal renal tubular acidosis and type II respiratory failure in rats

  2010cites this paper
• Cardiac Glycosides Induce Cell Death in Human Cells by Inhibiting General Protein Synthesis

  2009cites this paper
• Extracellular signal regulated kinase 5 mediates signals triggered by the novel tumor promoter palytoxin.

  2009cites this paper
• Up‐regulation of apical NHE3 in renal OK cells overexpressing the rodent alpha1‐subunit of the Na+ pump

  2006cites this paper
• Functional Characterization of Src-interacting Na/K-ATPase Using RNA Interference Assay*

  2006cites this paper
• Functional genomic dissection of multimeric protein families in zebrafish

  2003cites this paper
• Extrusion of intracellular calcium ion after in vitro ischemia in the rat hippocampal CA1 region.

  2002cites this paper
• Participation of Na,K-ATPase in FGF-2 secretion: rescue of ouabain-inhibitable FGF-2 secretion by ouabain-resistant Na,K-ATPase alpha subunits.

  2000cites this paper
• The Na,K-ATPase alpha4 gene (Atp1a4) encodes a ouabain-resistant alpha subunit and is tightly linked to the alpha2 gene (Atp1a2) on mouse chromosome 1.

  1999cites this paper
• Components of vectors for gene transfer and expression in mammalian cells.

  1999cites this paper
• Isozymes of the Na-K-ATPase: heterogeneity in structure, diversity in function.

  1998cites this paper
• Does binding of ouabain to human alpha1-subunit of Na+, K+-ATPase affect the ATPase activity of adjacent rat alpha1-subunit?

  1998cites this paper
• Identification of the Site of Inhibition by Omeprazole of a α-β Fusion Protein of the H,K-ATPase Using Site-directed Mutagenesis*

  1998cites this paper
• Expression of the rat liver Na+/taurocholate cotransporter is regulated in vivo by retention of biliary constituents but not their depletion

  1997cites this paper
• Regulation of rat Na(+)-K(+)-ATPase activity by PKC is modulated by state of phosphorylation of Ser-943 by PKA.

  1997cites this paper
• PKA-mediated phosphorylation and inhibition of Na(+)-K(+)-ATPase in response to beta-adrenergic hormone.

  1997cites this paper
• Regulation of rat Na+-K+-ATPase activity by PKC is modulated by state of phosphorylation of Ser-943 by PKA.

  1997cites this paper
• Localization of cytoplasmic and extracellular domains of Na,K-ATPase by epitope tag insertion.

  1996cites this paper
• Down-regulation of expression and function of the rat liver Na+/bile acid cotransporter in extrahepatic cholestasis.

  1996cites this paper
• Loss of the β1 subunit of the sodium pump during lymphocyte differentiation

  1996cites this paper
• Ouabain Interactions with the H5-H6 Hairpin of the Na,K-ATPase Reveal a Possible Inhibition Mechanism via the Cation Binding Domain*

  1996cites this paper
• Loss of the beta 1 subunit of the sodium pump during lymphocyte differentiation.

  1996cites this paper
• Chapter 4 Structure–Function Relationship of Na,K-ATPase: The Digitalis Receptor

  1994cites this paper
• Isoforms of the Na,K-ATPase: family members in search of function.

  1994cites this paper
• Ouabain-resistant OR6 cells express the murine alpha 1-subunit of the Na,K-ATPase with a T797-I797 substitution.

  1994cites this paper
• Aldosterone-mediated Na/K-ATPase expression is alpha 1 isoform specific in the renal cortical collecting duct.

  1993cites this paper
• A murine genomic DNA fragment amplifies ouabain-induced Na,K-ATPase alpha/beta-subunit mRNA up-regulation and confers ouabain resistance.

  1993cites this paper
• Transmembrane organization of the Na,K-ATPase determined by epitope addition.

  1993cites this paper
• Molecular cloning of a retina-specific membrane guanylyl cyclase.

  1992cites this paper
• Molecular cloning and characterization of Na,K-ATPase from Hydra vulgaris: implications for enzyme evolution and ouabain sensitivity.

  1992cites this paper
• Na+, K(+)-ATPase isoforms in the retina.

  1992cites this paper
• Molecular dissection of functional domains of the E1E2-ATPase using sodium and calcium pump chimeric molecules.

  1992cites this paper
• Mammalian expression vectors with modulatable promoters and two multiple cloning sites.

  1992cites this paper
• Interferon-alpha-induced gene expression: evidence for a selective effect of ouabain on activation of the ISGF3 transcription complex.

  1992cites this paper
• The functional role of the β‐subunit in the maturation and intracellular transport of Na,K‐ATPase

  1991cites this paper
• The sensitivity of insulin-stimulated and basal Na efflux to ouabain in frog skeletal muscle cells.

  1991cites this paper
• The functional role of the beta-subunit in the maturation and intracellular transport of Na,K-ATPase.

  1991cites this paper
• Abundance of Na(+)-K(+)-ATPase mRNA is regulated by glucocorticoid hormones in infant rat kidneys.

  1991cites this paper
• Diabetes Mellitus and Glucagon Alter Ouabain-Sensitive Na+-K+-ATPase in Rat Small Intestine

  1991cites this paper
• Ouabain‐ and Ca2+‐sensitive ATPase activity of chimeric Na‐ and Ca‐pump molecules

  1991cites this paper
• Structure-function relationship of Na,K-ATPase.

  1991cites this paper
• Subunit assembly and functional maturation of Na,K-ATPase

  1990cites this paper
• Ouabain-resistant mutants of the rat Na,K-ATPase alpha 2 isoform identified by using an episomal expression vector

  1990cites this paper
• Reconstitution and fractionation of renal brush border transport proteins.

  1990cites this paper
• Differential expression and enzymatic properties of the Na+,K(+)-ATPase alpha 3 isoenzyme in rat pineal glands.

  1990cites this paper
• Giant MDCK cells: a novel expression system.

  1990cites this paper
• Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments

  1990cites this paper
• Isozymes of the Na+/K+-ATPase.

  1989cites this paper
• The search for the endogenous digitalis: an alternative hypothesis.

  1989cites this paper
• Identification of a region within the Na,K-ATPase alpha subunit that contributes to differential ouabain sensitivity

  1989cites this paper
• Antisera specific for the alpha 1, alpha 2, alpha 3, and beta subunits of the Na,K-ATPase: differential expression of alpha and beta subunits in rat tissue membranes.

  1989cites this paper
• Antisera specific for the .alpha.1, .alpha.2, .alpha.3, and .beta. subunits of the sodium-potassium ATPase: differential expression of .alpha. and .beta. subunits in rat tissue membranes

  1989cites this paper
• Expression of sodium pump activities in BALB/c 3T3 cells transfected with cDNA encoding α3‐subunits of rat brain Na+,K+‐ATPase

  1988cites this paper