title: Just one position-independent lysine residue can direct MelanA into proteasomal degradation following N-terminal fusion of ubiquitin.
abstract: N-terminal stable in frame fusion of ubiquitin (Ub) has been shown to target the fusion protein for proteasomal degradation. This pathway, called the Ub fusion degradation (UFD), might also elevate MHC class I (MHC-I) antigen presentation of specific antigens. The UFD, mainly studied on cytosolic proteins, has been described to be mediated by polyubiquitination of specific lysine residues within the fused Ub moiety. Using the well characterized melanoma-specific antigen MelanA as a model protein, we analyzed the requirements of the UFD for ubiquitination and proteasomal degradation of a transmembrane protein. Here we show that fusion of the non-cleavable Ub(G76V) variant to the N-terminus of MelanA results in rapid proteasomal degradation via the endoplasmic reticulum-associated degradation (ERAD) pathway and, consequently, leads to an increased MHC-I antigen presentation. While lysine residues within Ub are dispensable for these effects, the presence of one single lysine residue, irrespectively of its location along the fusion protein, is sufficient to induce degradation of MelanA. These results show that the ubiquitination, ER to cytosol relocation and proteasomal degradation of a transmembrane protein can be increased by N-terminal fusion of Ub at the presence of at least one, position independent lysine residue. These findings are in contrast to the conventional wisdom concerning the UFD and indicate a new concept to target a protein into the ubiquitin-proteasome system (UPS) and thus for enhanced MHC-I antigen presentation, and might open up new possibilities in the development of tumor vaccines.
authors: Institute of Clinical and Molecular Virology, Universitätsklinikum Erlangen, Erlangen, Germany.
authors: Christian
authors: C
authors: Setz
authors: Melanie
authors: M
authors: Friedrich
authors: Sabine
authors: S
authors: Hahn
authors: Jan
authors: J
authors: Dörrie
authors: Niels
authors: N
authors: Schaft
authors: Gerold
authors: G
authors: Schuler
authors: Ulrich
authors: U
authors: Schubert
mesh_terms: D017951:Antigen Presentation / Q000235:genetics / Q000502:physiology; D015153:Blotting, Western; D002460:Cell Line; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D058950:Melanoma-Specific Antigens / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D058950:Melanoma-Specific Antigens; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex; D008239:Lysine
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