Identification of RNA-binding proteins in macrophages by interactome capture
Biochemistry
Cell Biology
Final Report Abstract
Macrophages exert the primary cellular innate immune response. Pathogen components like bacterial lipopolysaccharides (LPS) initiate Toll-like receptor 4 (TLR4) signaling, which involves mitogen activated protein kinases (MAPK) and nuclear factor kappa B (NFκB) in different pathway branches and ultimately induces inflammatory cytokine and chemokine expression, macrophage migration and phagocytosis. Importantly, post-transcriptional control of TLR4 downstream signaling contributes to the tight regulation of inflammatory cytokine synthesis. Emerging evidence highlights the role of RNA binding proteins (RBPs) in the post-transcriptional control of innate immune response. Previously, we identified the poly(A)+ RNA interactome of RAW 264.7 cells. Of 402 RBPs, 32 were classified as unique in macrophages, including nineteen not reported to interact with nucleic acids before. Remarkably, P23 a HSP90 co-chaperone, also known as cytosolic prostaglandin E2 synthase (PTGES3), exhibited differential poly(A)+ RNA binding in untreated and LPS-induced macrophages. To identify mRNAs bound by P23 and to elucidate potential regulatory RBP functions, we immunoprecipitated P23 from cytoplasmic extracts of cross-linked untreated and LPS-induced macrophages. RNAseq revealed reduced binding of 44 mRNAs in response to LPS. Kif15 mRNA, which encodes kinesin family member 15 (KIF15), a motor protein implicated in cytoskeletal reorganization and cell mobility was selected for further analysis. Noteworthy, phagocytic activity of LPS-induced macrophages was enhanced by P23 depletion. Specifically, in untreated RAW 264.7 macrophages, decreased P23 results in Kif15 mRNA destabilization, diminished KIF15 expression and accelerated macrophage migration. We show that the unexpected RBP function of P23 contributes to the regulation of macrophage phagocytotic activity and migration.
Publications
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Translation control of TAK1 mRNA by hnRNP K modulates LPS-induced macrophage activation. RNA, 20(6), 899-911.
Liepelt, Anke; Mossanen, Jana C.; Denecke, Bernd; Heymann, Felix; De Santis, Rebecca; Tacke, Frank; Marx, Gernot; Ostareck, Dirk H. & Ostareck-Lederer, Antje
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Identification of RNA-binding Proteins in Macrophages by Interactome Capture. Molecular & Cellular Proteomics, 15(8), 2699-2714.
Liepelt, Anke; Naarmann-de Vries, Isabel S.; Simons, Nadine; Eichelbaum, Katrin; Föhr, Sophia; Archer, Stuart K.; Castello, Alfredo; Usadel, Björn; Krijgsveld, Jeroen; Preiss, Thomas; Marx, Gernot; Hentze, Matthias W.; Ostareck, Dirk H. & Ostareck-Lederer, Antje
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US 9745369 B2 08/29/2017, EP 2855519 B1 11/15/2017: Modulation of the TLR4-signaling pathway
Ostareck-Lederer, A.; Ostareck, D.H.; Marx, G.; Liepelt A. & Mossanen, J.
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RNA-Binding Proteins in the Control of LPS-Induced Macrophage Response. Frontiers in Genetics, 10(c(2019, 2, 4)).
Ostareck, Dirk H. & Ostareck-Lederer, Antje
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Deep and accurate detection of m6A RNA modifications using miCLIP2 and m6Aboost machine learning. Nucleic Acids Research, 49(16), e92-e92.
Körtel, Nadine; Rücklé, Cornelia; Zhou, You; Busch, Anke; Hoch-Kraft, Peter; Sutandy, F X Reymond; Haase, Jacob; Pradhan, Mihika; Musheev, Michael; Ostareck, Dirk; Ostareck-Lederer, Antje; Dieterich, Christoph; Hüttelmaier, Stefan; Niehrs, Christof; Rausch, Oliver; Dominissini, Dan; König, Julian & Zarnack, Kathi
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Methylated HNRNPK acts on RPS19 to regulate ALOX15 synthesis in erythropoiesis. Nucleic Acids Research, 49(6), 3507-3523.
Naarmann-de Vries, Isabel S; Senatore, Roberta; Moritz, Bodo; Marx, Gernot; Urlaub, Henning; Niessing, Dierk; Ostareck, Dirk H & Ostareck-Lederer, Antje
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P23 Acts as Functional RBP in the Macrophage Inflammation Response. Frontiers in Molecular Biosciences, 8(c(2021, 6, 11)).
de Vries, Sebastian; Benes, Vladimir; Naarmann-de Vries, Isabel S.; Rücklé, Cornelia; Zarnack, Katharina; Marx, Gernot; Ostareck, Dirk H. & Ostareck-Lederer, Antje
