A novel C53/LZAP-interacting protein regulates stability of C53/LZAP and DDRGK domain-containing Protein 1 (DDRGK1) and modulates NF-kappaB signaling.
C53/LZAP (also named as Cdk5rap3) is a putative tumor suppressor that plays important roles in multiple cell signaling pathways, including DNA damage response and NF-kappaB signaling. Yet how its function is regulated remains largely unclear. Here we report the isolation and characterization of two novel C53/LZAP-interacting proteins, RCAD (Regulator of ... C53/LZAP and DDRGK1) and DDRGK1 (DDRGK domain-containing protein 1). Our co-immunoprecipitation assays confirmed their interactions, while gel filtration assay indicated that C53/LZAP and RCAD may form a large protein complex. Intriguingly, we found that RCAD knockdown led to dramatic reduction of C53/LZAP and DDRGK1 proteins. We also found that C53/LZAP and DDRGK1 became more susceptible to the proteasome-mediated degradation in RCAD knockdown cells, whereas their ubiquitination was significantly attenuated by RCAD overexpression. In addition, we found that RCAD, like C53/LZAP, also plays an important role in regulation of NF-kappaB signaling and cell invasion. Taken together, our findings strongly suggest that RCAD is a novel regulator of C53/LZAP tumor suppressor and NF-kappaB signaling.
Mesh Terms:
Base Sequence, Blotting, Northern, Cell Line, Tumor, Chromatography, Gel, DNA Primers, Gene Knockdown Techniques, Humans, Intracellular Signaling Peptides and Proteins, Microscopy, Electron, NF-kappa B, Nerve Tissue Proteins, Polymerase Chain Reaction, Signal Transduction
Base Sequence, Blotting, Northern, Cell Line, Tumor, Chromatography, Gel, DNA Primers, Gene Knockdown Techniques, Humans, Intracellular Signaling Peptides and Proteins, Microscopy, Electron, NF-kappa B, Nerve Tissue Proteins, Polymerase Chain Reaction, Signal Transduction
J. Biol. Chem.
Date: May. 14, 2010
PubMed ID: 20228063
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