Premium
NEDD4 mediated degradation of target proteins under simulated microgravity conditions
Author(s) -
Fiedorowicz Katarzyna,
Da SilvaAzevedo Luis,
Kuebler Wolfgang Michael,
Zakrzewicz Andreas,
Pries Axel Radlach
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.1035.4
Subject(s) - downregulation and upregulation , umbilical vein , nedd4 , ubiquitin ligase , ubiquitin , chemistry , microbiology and biotechnology , protein degradation , shear stress , blot , biophysics , biology , biochemistry , materials science , in vitro , gene , composite material
The effects of microgravity on human vascular endothelial cells may contribute to altered cardiovascular functions in space. Since endothelial function critically depends on protein synthesis and degradation, we addressed the effects of microgravity on the ubiquitin‐ligase NEDD4 in endothelial cells. Human umbilical vein endothelial cells or EAhy926 cells were exposed to simulated microgravity in the Rotating Wall Vessel (RWV) for 24 h, 48 h and 72 h. Controls were kept under normal gravity or exposed to shear stress using a cone and plate apparatus (6 dyn\cm 2 , 24 h). Gene expression and protein concentration were analysed by real‐time RT‐PCR and Western blotting, respectively. Our results show a two fold upregulation of NEDD4 under microgravity and no regulation by shear stress (relative to controls 1G, no shear). NEDD4 targets e.g. Cx43, Notch1, eps15, IGF1R, PLCgamma1 were strongly downregulated after 48 h and 72 h exposure to microgravity. These data suggest that NEDD4 expression is sensitive to gravity, which may be a mechanism to enhance protein degradation in endothelial cells.