title: TP53 regulates miRNA association with AGO2 to remodel the miRNA-mRNA interaction network.
abstract: DNA damage activates TP53-regulated surveillance mechanisms that are crucial in suppressing tumorigenesis. TP53 orchestrates these responses directly by transcriptionally modulating genes, including microRNAs (miRNAs), and by regulating miRNA biogenesis through interacting with the DROSHA complex. However, whether the association between miRNAs and AGO2 is regulated following DNA damage is not yet known. Here, we show that, following DNA damage, TP53 interacts with AGO2 to induce or reduce AGO2's association of a subset of miRNAs, including multiple let-7 family members. Furthermore, we show that specific mutations in TP53 decrease rather than increase the association of let-7 family miRNAs, reducing their activity without preventing TP53 from interacting with AGO2. This is consistent with the oncogenic properties of these mutants. Using AGO2 RIP-seq and PAR-CLIP-seq, we show that the DNA damage-induced increase in binding of let-7 family members to the RISC complex is functional. We unambiguously determine the global miRNA-mRNA interaction networks involved in the DNA damage response, validating them through the identification of miRNA-target chimeras formed by endogenous ligation reactions. We find that the target complementary region of the let-7 seed tends to have highly fixed positions and more variable ones. Additionally, we observe that miRNAs, whose cellular abundance or differential association with AGO2 is regulated by TP53, are involved in an intricate network of regulatory feedback and feedforward circuits. TP53-mediated regulation of AGO2-miRNA interaction represents a new mechanism of miRNA regulation in carcinogenesis.
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Jonathan
authors.initials: J
authors.lastname: Krell
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Justin
authors.initials: J
authors.lastname: Stebbing
authors.affiliation: Department of Cellular Biotechnologies and Hematology, Division of Molecular Genetics, "Sapienza" University of Rome, Rome 00161, Italy;
authors.forename: Claudia
authors.initials: C
authors.lastname: Carissimi
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Aleksandra F
authors.initials: AF
authors.lastname: Dabrowska
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Alexander
authors.initials: A
authors.lastname: de Giorgio
authors.affiliation: Department of Surgery and Cancer, HPB Surgical Unit, Imperial College, Hammersmith Hospital Campus, London W12 0NN, United Kingdom;
authors.forename: Adam E
authors.initials: AE
authors.lastname: Frampton
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Victoria
authors.initials: V
authors.lastname: Harding
authors.affiliation: Department of Cellular Biotechnologies and Hematology, Division of Molecular Genetics, "Sapienza" University of Rome, Rome 00161, Italy;
authors.forename: Valerio
authors.initials: V
authors.lastname: Fulci
authors.affiliation: Department of Cellular Biotechnologies and Hematology, Division of Molecular Genetics, "Sapienza" University of Rome, Rome 00161, Italy;
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Macino
authors.affiliation: Institute for System Analysis and Computer Science "Antonio Ruberti," National Research Council, Rome 00185, Italy; Department of Biotechnology, University of Verona, Verona 37134, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Colombo
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine, London W12 0NN, United Kingdom;
authors.forename: Leandro
authors.initials: L
authors.lastname: Castellano
mesh_terms: D060565:Argonaute Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D055786:Gene Knockout Techniques; D053263:Gene Regulatory Networks; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D011485:Protein Binding; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics*; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin