Article

The sarcolemmal calcium pump inhibits the calcineurin/nuclear factor of activated T-cell pathway via interaction with the calcineurin A catalytic subunit

The University of Manchester, Manchester, England, United Kingdom
Journal of Biological Chemistry (Impact Factor: 4.6). 09/2005; 280(33):29479-87. DOI: 10.1074/jbc.M501326200
Source: PubMed

ABSTRACT The calcineurin/nuclear factor of activated T-cell (NFAT) pathway represents a crucial transducer of cellular function. There is increasing evidence placing the sarcolemmal calcium pump, or plasma membrane calcium/calmodulin ATPase pump (PMCA), as a potential modulator of signal transduction pathways. We demonstrate a novel interaction between PMCA and the calcium/calmodulin-dependent phosphatase, calcineurin, in mammalian cells. The interaction domains were located to the catalytic domain of PMCA4b and the catalytic domain of the calcineurin A subunit. Endogenous calcineurin activity, assessed by measuring the transcriptional activity of its best characterized substrate, NFAT, was significantly inhibited by 60% in the presence of ectopic PMCA4b. This inhibition was notably reversed by the co-expression of the PMCA4b interaction domain, demonstrating the functional significance of this interaction. PMCA4b was, however, unable to confer its inhibitory effect in the presence of a calcium/calmodulin-independent constitutively active mutant calcineurin A suggesting a calcium/calmodulin-dependent mechanism. The modulatory function of PMCA4b is further supported by the observation that endogenous calcineurin moves from the cytoplasm to the plasma membrane when PMCA4b is overexpressed. We suggest recruitment by PMCA4b of calcineurin to a low calcium environment as a possible explanation for these findings. In summary, our results offer strong evidence for a novel functional interaction between PMCA and calcineurin, suggesting a role for PMCA as a negative modulator of calcineurin-mediated signaling pathways in mammalian cells. This study reinforces the emerging role of PMCA as a molecular organizer and regulator of signaling transduction pathways.

0 Followers
 · 
67 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: The mechanisms linking the expression of sarcomeric mutant proteins to the development of pathological hypertrophy in hypertrophic cardiomyopathy (HCM) remain poorly understood. We investigated the role of the plasma membrane Ca(2+)-ATPase PMCA4 in the HCM phenotype using a transgenic model that expresses mutant (Glu180Gly) α-tropomyosin (Tm180) in heart. Immunoblot analysis revealed that cardiac PMCA4 expression was upregulated early in Tm180 disease pathogenesis. This was accompanied by an increase in levels of the L-type Ca(2+)-channel, which is implicated in pathological hypertrophy. When Tm180 mice were crossed with a PMCA4-null line, loss of PMCA4 caused the abrogation of hypertrophy in Tm180/PMCA4-null double mutant mice. RT-PCR analysis of Tm180/PMCA4-null hearts revealed blunting of the fetal program and reversion of pro-fibrotic Col1a1 and Col3a1 gene expression to wild-type levels. This was accompanied by evidence of reduced L-type Ca(2+)-channel expression, and diminished calcineurin activity. Expression of the metabolic substrate transporters glucose transporter 4 and carnitine palmitoyltransferase 1b was preserved and Tm180-related changes in mRNA levels of various contractile stress-related proteins including the cardiac ankyrin protein CARP and the N2B isoform of titin were reversed in Tm180/PMCA4-null hearts. cGMP levels were increased and phosphorylation of vasodilator-stimulated phosphoprotein was elevated in Tm180/PMCA4-null hearts. These changes were associated with a sharp reduction in left ventricular end-diastolic pressure in Tm180/PMCA4-null hearts, which occurred despite persistence of Tm180-related impairment of relaxation dynamics. These results reveal a novel and specific role for PMCA4 in the Tm180 hypertrophic phenotype, with the "protective" effects of PMCA4 deficiency encompassing multiple determinants of HCM-related hypertrophy.
    Journal of Molecular and Cellular Cardiology 09/2014; 77. DOI:10.1016/j.yjmcc.2014.09.025 · 5.22 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Vascular endothelial growth factor (VEGF) has been identified as a crucial regulator of physiological and pathological angiogenesis. Among the intracellular signaling pathways triggered by VEGF, activation of the calcineurin/nuclear factor of activated T cells (NFAT) signaling axis has emerged as a critical mediator of angiogenic processes. We and others previously reported a novel role for the plasma membrane calcium ATPase (PMCA) as an endogenous inhibitor of the calcineurin/NFAT pathway, via interaction with calcineurin, in cardiomyocytes and breast cancer cells. However, the functional significance of the PMCA/calcineurin interaction in endothelial pathophysiology has not been addressed thus far.
    Arteriosclerosis Thrombosis and Vascular Biology 08/2014; 34(10). DOI:10.1161/ATVBAHA.114.304363 · 5.53 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Plant Ca2+ATPases regulate many signalling pathways which are important for plant growth, development and abiotic stress responses. Our previous work identified that overexpression of OsACA6 confers salinity and drought tolerance in tobacco. In the present work we report, the function of OsACA6 in cold stress tolerance in transgenic tobacco plants. The expression of OsACA6 was induced by cold stress. The promoter-GUS fusion analyses in the different tissues of transgenic tobacoco confirmed that OsACA6 promoter is cold stress-inducible. Transgenic tobacco plants overexpressing OsACA6 exhibited cold tolerance compared to the wild type (WT) controls. The enhanced tolerance was confirmed by phenotypic analyses as well as by measuring germination, survival rate, chlorophyll content, cell membrane stability, malondialdehyde and proline content. Compared to the WT, the expression of catalase, ascorbate peroxidase and superoxide dismutase increased in the OsACA6 overexpressing plants, which was inversely correlated with the levels of H2O2 in the transgenic lines. We also identified interacting proteins of OsACA6 by using yeast two-hybrid screening assay. Most of the interacting partners of OsACA6 are associated with the widespread biological processes including plant growth, development, signalling and stress adaptation. Furthermore, we also confirmed that OsACA6 is able to self-interact. Overall, these results suggest that OsACA6 plays an important role in cold tolerance at least in part, by regulating antioxidants-mediated removal of reactive oxygen species or by interacting with different calcium signal decoders including calmodulin-like proteins (CaM) calcium/calmodulin dependent protein kinases (CDPKs) and receptor-like protein kinases (RLKs).
    Plant Physiology and Biochemistry 06/2014; 82. DOI:10.1016/j.plaphy.2014.06.007 · 2.35 Impact Factor