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The Regulation of Cellular Proliferation by VACM‐1/CUL5 is Dependent on its Post‐translational Modifications by NEDD8
Author(s) -
Lee Si Eun,
Sundquist Skylar,
BurnatowskaHledin Maria
Publication year - 2018
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.2018.32.1_supplement.791.15
Subject(s) - cullin , ubiquitin ligase , biology , microbiology and biotechnology , in vitro , nedd8 , ubiquitin , biochemistry , gene
VACM‐1/CUL5 is a member of the cullin family of proteins involved in the E3 ligase complex formation and subsequent protein degradation by ubiquitin‐dependent pathways. In vivo , VACM‐1/CUL5 is expressed in endothelial cells, and when expressed in vitro it attenuates cellular growth. While the specific mechanism by which VACM‐1/CUL5 suppresses cell proliferation is unknown, its antiproliferative effect is dependent on its post‐translational modification by an ubiquitin‐like protein, NEDD8. To investigate the effect of NEDD8 on the structure and function of VACM‐1/CUL5, we have mutated its cDNA sequence at the putative neddylation sites and expressed the mutated construct in Rat Adrenal Medulla Endothelial Cells (RAMEC) and COS‐1 cells in vitro. Our results demonstrate that the antiproliferative effect of wt VACM/CUL5 was attenuated in cells transfected with the mutated VACM‐1/CUL5 cDNA. Furthermore, the mutated VACM‐1/CUL5 affected the anti‐angiogenic phenotype of wt VACM‐1 when expressed in RAMEC cells and grown on Matrigel® support. Finally, the immunoblotting results show that the mutant is modified at multiple sites by NEDD8 in vitro . These results suggest that antiproliferative effect of VACM‐1/CUL5 is regulated by its neddylation status. Support or Funding Information This work is supported by the Paul A. Schaap Fellowship and Hope College Department of Biology. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .