The alternative translated MDMXp60 isoform regulates MDM2 activity
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Isoforms derived from alternative splicing, mRNA translation initiation or promoter usage extend the functional repertoire of the p53, p63 and p73 genes family and of their regulators MDM2 and MDMX. Here we show capindependent translation of an N-terminal truncated isoform of hMDMX, hMDMXp60, which is initiated at the 7th AUG codon downstream of the initiation site for full length hMDMXFL at position C384. hMDMXp60 lacks the p53 binding motif but retains the RING domain and interacts with hMDM2 and hMDMXFL. hMDMXp60 shows higher affinity for hMDM2, as compared to hMDMXFL. In vitro data reveal a positive cooperative interaction between hMDMXp60 and hMDM2 and in cellulo data show that low levels of hMDMXp60 promote degradation of hMDM2 whereas higher levels stabilize hMDM2 and prevent hMDM2-mediated degradation of hMDMXFL. These results describe a novel alternatively translated hMDMX isoform that exhibits unique regulatory activity toward hMDM2 autoubiquitination. The data illustrate how the N-terminus of hMDMX regulates its C-terminal RING domain and the hMDM2 activity. © 2015 Taylor %26 Francis Group, LLC.
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Alternative translation; Isoforms; MDM2; MDMX; Protein stability cell DNA; complementary DNA; messenger RNA; polyclonal antibody; protein MDM2; protein MDMX; protein p53; protein p60; ubiquitin protein ligase E3; capped RNA; codon; isoprotein; MDM2 protein, human; MDM4 protein, human; messenger RNA; nuclear protein; oncoprotein; protein binding; protein MDM2; 5' untranslated region; Article; cell lysate; codon; controlled study; enzyme activity; enzyme linked immunosorbent assay; human; human cell; immunoprecipitation; in vitro study; open reading frame; oscillation; protein degradation; protein expression; protein stability; reverse transcription polymerase chain reaction; RING finger motif; translation initiation; ubiquitination; ultraviolet irradiation; Western blotting; alternative RNA splicing; cell line; genetics; metabolism; protein synthesis; Alternative Splicing; Cell Line; Codon; Humans; Nuclear Proteins; Protein Binding; Protein Biosynthesis; Protein Isoforms; Protein Stability; Proteolysis; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-mdm2; RNA Caps; RNA, Messenger
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