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Hoefig, Kai P.; Reim, Alexander; Gallus, Christian; Wong, Elaine H.; Behrens, Gesine; Conrad, Christine; Xu, Meng; Kifinger, Lisa; Ito-Kureha, Taku; Defourny, Kyra A. Y.; Geerlof, Arie; Mautner, Josef; Hauck, Stefanie M.; Baumjohann, Dirk; Feederle, Regina; Mann, Matthias; Wierer, Michael; Glasmacher, Elke and Heissmeyer, Vigo (2021): Defining the RBPome of primary T helper cells to elucidate higher-order Roquin-mediated mRNA regulation. In: Nature Communications, Vol. 12, No. 1, 5208

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Post-transcriptional gene regulation in T cells is dynamic and complex as targeted transcripts respond to various factors. This is evident for the Icos mRNA encoding an essential costi-mulatory receptor that is regulated by several RNA-binding proteins (RBP), including Roquin-1 and Roquin-2. Here, we identify a core RBPome of 798 mouse and 801 human T cell proteins by utilizing global RNA interactome capture (RNA-IC) and orthogonal organic phase separation (OOPS). The RBPome includes Stat1, Stat4 and Vav1 proteins suggesting unexpected functions for these transcription factors and signal transducers. Based on proximity to Roquin-1, we select similar to 50 RBPs for testing coregulation of Roquin-1/2 targets by induced expression in wild-type or Roquin-1/2-deficient T cells. Besides Roquin-independent contributions from Rbms1 and Cpeb4 we also show Roquin-1/2-dependent and target-specific coregulation of Icos by Celf1 and Igf2bp3. Connecting the cellular RBPome in a post-transcriptional context, we find contributions from multiple RBPs to the prototypic regulation of mRNA targets by individual trans-acting factors.

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