Mass Spec Molecular Oncology Charpentier et al., 2021

These files contain the full analysed mass spec data of the results presented in Charpentier et al., 2021 Molecular Oncology.
We focused on MELOE-1, a melanoma neoantigen whose expression is restricted to tumor cells as a result of lineage-specific transcription and tumor-specific IRES-dependent translation of the polycistronic meloe RNA (Godet et al., 2008 and Bobinet et al., 2013).
Our previous study on MELOE-1 translation reported the presence of an IRES activity located around 250nt upstream of MELOE-1 ORF (Carbonnelle et al., 2013). In the present work, we looked for the ITAF(s) that could bind to this region. To this aim, we in vitro transcribed the 275nt RNA sequence upstream of MELOE-1 ORF, biotinylated it and coupled it to a streptavidin BIAcore chip. We then prepared whole cell lysates from three melanoma cell-lines M117, M134 and M170, ran them on the RNA-coated BIAcore chip (40 rounds of 5 min injections) and then recovered the eluted material for mass spectrometry analysis.
references:
Godet Y et al. MELOE-1 is a new antigen overexpressed in melanomas and involved in adoptive T cell transfer efficiency. J Exp Med 2008;205(11):2673–2682.
Bobinet M et al. Overexpression of Meloe Gene in Melanomas Is Controlled Both by Specific Transcription Factors and Hypomethylation. PLoS ONE 2013;8(9):e75421.
Carbonnelle D et al. The Melanoma Antigens MELOE-1 and MELOE-2 Are Translated from a Bona Fide Polycistronic mRNA Containing Functional IRES Sequences. PLoS ONE 2013;8(9).