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Shared sequence features of diverse oncofusions promote gain-of-function RNA Pol II partitioning to drive gene activation

ABSTRACT: Translocation renal cell carcinoma is a rare but aggressive cancer driven by oncogenic fusions between the DNA-binding transcription factor TFE3 and diverse subunits. We have found that these diverse fusions promote condensate formation and activation of target genes. To investigate what the condensates formed by different TFE3 fusions partition we used a cell-free method relying on condensate reconstitution within a nuclear extract. Using recombinant purified PRCC-TFE3 or ASPL-TFE3, we reconstituted condensates in a soluble nuclear extract. The extract was then fractionated by centrifugation into a supernatant and pellet. For each fusion supernatant and pellet fractions were collected in triplicate. Samples were individually labeled using a 6-plex tandem mass tag (TMT) post-tryptic isobaric labeling strategy and the three supernatant and three pellet fractions of each fusion were mixed and analyzed by LC-MS to provide quantitative measurements of partitioning. In summary, we deposit 2 TMT 6-plex samples which represent triplicates for each PRCC-TFE3 (1071162 F1-8) and ASPL-TFE3 (1069084 F1-8). TMT labels 126, 127, 128: Sup TMT labels 129, 130, 131: Pellet

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Benjamin Sabari

PROVIDER: MSV000094715 | MassIVE | Wed May 08 08:38:00 BST 2024

REPOSITORIES: MassIVE

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Shared sequence features of diverse oncofusions promote gain-of-function RNA Pol II partitioning to drive gene activation

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