(Verbatim from the application): Both acute and chronic regulation of the Na,K-ATPase by prostaglandins will be examined using the Madin Darby Canine Kidney (MDCK) cell line in hormonally defined serum free medium. Two classes of variant MDCK cells will be utilized, includingProstaglandin El (PGE1) independent MDCK cells (with elevated cyclic AMP (cAMP)), which no longer require PGE1 for growth, and dibutyryl cAMP resistant MDCK cells (with a defect in CAMP dependent protein kinase (PKA), which no longer respond to PGE1 by an increase Na,K-ATPase activity). The hypothesis will be evaluated that following long term PGE1 treatment, the Na,K-ATPase is regulated at the transcriptional level via the protein kinase C (PKC) as well as the PKA pathways, unlike the case with a short term PGE1 treatment, in which the Na,K-ATPase is regulated post-translationally. In order to evaluate this hypothesis: (1) Transcriptional regulation of the Na,K-ATPase beta subunit gene will be examined by means of transient transfection studies with normal and variant MDCK cells, utilizing the pH beta1-1141 Luc plasmid. The role of the PKA, PKC and calcium pathways will be examined. Regulatory regions in the Na,K-ATPase beta subunit promoter are to be defined, and trans-acting factors identified. (2) To determine the effect of chronic PGE1 treatment on transcriptional regulation of endogenous genes, nuclear-runoff transcription studies will be conducted with normal and variant MDCK cells. Control studies will also be conducted with primary rabbit kidney proximal tubule cells. Post-transcriptional effects of PGE1 will be evaluated by means of Northern analysis, an assessment of effects on mRNA stability. Na,K-ATPase biosynthesis and degradation rates will be determined. (3) To determine the mechanism underlying acute effects of PGE1, PGE2 and 8Br-cAMP, the possibility will be evaluated that the Na,K-ATPase is phosphorylated by these agents. The possible involvement of an inhibitory phosphatase, in regulating the phosphorylation of the Na,K-ATPase will also be examined.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL069676-01
Application #
6285702
Study Section
Cardiovascular and Renal Study Section (CVB)
Program Officer
Barouch, Winifred
Project Start
2001-06-10
Project End
2005-05-31
Budget Start
2001-06-10
Budget End
2002-05-31
Support Year
1
Fiscal Year
2001
Total Cost
$271,104
Indirect Cost
Name
State University of New York at Buffalo
Department
Biochemistry
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Taub, Mary; Cutuli, Facundo (2016) Data on Na,K-ATPase in primary cultures of renal proximal tubule cells treated with catecholamines. Data Brief 6:419-22
Saidani, Chanez; Hammoudi-Triki, Djelila; Laraba-Djebari, Fatima et al. (2016) In vitro studies with renal proximal tubule cells show direct cytotoxicity of Androctonus australis hector scorpion venom triggered by oxidative stress, caspase activation and apoptosis. Toxicon 120:29-37
Taub, Mary; Garimella, Sudha; Kim, Dongwook et al. (2015) Renal proximal tubule Na,K-ATPase is controlled by CREB-regulated transcriptional coactivators as well as salt-inducible kinase 1. Cell Signal 27:2568-78
Taub, Mary; Springate, James E; Cutuli, Facundo (2010) Targeting of renal proximal tubule Na,K-ATPase by salt-inducible kinase. Biochem Biophys Res Commun 393:339-44
Matlhagela, Keikantse; Taub, Mary (2006) Prostaglandins regulate transcription by means of prostaglandin response elements located in the promoters of mammalian Na,K-ATPase beta 1 subunit genes. Ann N Y Acad Sci 1091:233-43
Borsick, Maryanne; Rajkhowa, Trivikram; Taub, Mary (2006) Evidence for post-transcriptional regulation of Na,K-ATPase by prostaglandin E1. Biochem Biophys Res Commun 345:739-45
Matlhagela, Keikantse; Taub, Mary (2006) Involvement of EP1 and EP2 receptors in the regulation of the Na,K-ATPase by prostaglandins in MDCK cells. Prostaglandins Other Lipid Mediat 79:101-13
Matlhagela, Keikantse; Taub, Mary (2006) Regulation of the Na-K-ATPase beta(1)-subunit promoter by multiple prostaglandin-responsive elements. Am J Physiol Renal Physiol 291:F635-46
Taub, Mary (2005) Primary kidney proximal tubule cells. Methods Mol Biol 290:231-47
Han, Ho Jae; Lee, Yun Jung; Park, Su Hyung et al. (2005) High glucose-induced oxidative stress inhibits Na+/glucose cotransporter activity in renal proximal tubule cells. Am J Physiol Renal Physiol 288:F988-96

Showing the most recent 10 out of 15 publications