microRNAs (miRNAs) are an abundant class of small RNAs that derive from distinctive hairpin precursor transcripts. Their biogenesis typically involves cleavage by the Drosha and Dicer RNase III enzymes, and the resultant mature miRNAs mediate extensive networks of post-transcriptional regulation. In the course of analyzing the miRNA pathway in Drosophila, we uncovered atypical miRNAs derived from short hairpin introns that we term mirtrons. Their biogenesis substitutes a seemingly obligate processing event by the Drosha RNase III enzyme with the splicing machinery. We characterized the biogenesis and evolutionary range of mirtrons across four animal species, using biochemical, molecular, genetic, and computational methods. This work led to a series of unexpected avenues that further expanded the diversity of non- canonical miRNAs, including a hybrid pathway that marries splicing with the RNA exosome to generate hairpins that serve as Dicer substrates, as well as short hairpins that generate miRNAs in a Dicer-independent mechanism in vertebrates. In addition, this work served as a foundation of our biological studies of miRNA functions. For our renewal application, we selected several directions on understanding enzymes and mechanisms that mediate the biogenesis of mirtrons and Dicer-independent miRNAs. We present extensive unpublished data for our proposed Aims, including the discovery of hundreds of mammalian mirtrons, including a novel 3'tailed subtype in mammals, the first demonstration of Dicer-independent miRNA function in Drosophila, and establishment of in vitro assays for Dicer-independent miRNA production. These preliminary data are the basis of individual gene studies to understand detailed mechanisms, which we will extend with genomewide perspectives. In addition, we wish to relate these findings to their broader implications with respect to other small RNAs (i.e. canonical miRNAs) and discovery of novel substrates. Finally, we are concerned with relating our research to larger evolutionary questions, including the evolutionary emergence of miRNAs and of RNAi itself, and with exploiting our mechanistic knowledge to advance RNA silencing as a technological tool.

Public Health Relevance

microRNAs are an extensive class of ~22 nucleotide RNAs that control the activity of messenger RNAs, the templates for protein synthesis. microRNAs dysfunction is linked to disease and cancer, while reciprocally, there is great potential for exploiting microRNAs and related regulatory RNAs as research tools and therapeutic strategies. This proposal extends our long-standing commitment to dissecting mechanisms of microRNA biogenesis by exploring new pathways that generate microRNAs and investigating enzymes that influence microRNA function.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
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Molecular Genetics B Study Section (MGB)
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Bender, Michael T
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Sloan-Kettering Institute for Cancer Research
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Kavaler, Joshua; Duan, Hong; Aradhya, Rajaguru et al. (2018) miRNA suppression of a Notch repressor directs non-neuronal fate in Drosophila mechanosensory organs. J Cell Biol 217:571-583
Duan, Hong; de Navas, Luis F; Hu, Fuqu et al. (2018) The mir-279/996 cluster represses receptor tyrosine kinase signaling to determine cell fates in the Drosophila eye. Development 145:
Zhou, Li; Lim, Mandy Yu Theng; Kaur, Prameet et al. (2018) Importance of miRNA stability and alternative primary miRNA isoforms in gene regulation during Drosophila development. Elife 7:
Teijeiro, Virginia; Yang, Dapeng; Majumdar, Sonali et al. (2018) DICER1 Is Essential for Self-Renewal of Human Embryonic Stem Cells. Stem Cell Reports 11:616-625
Mohammed, Jaaved; Flynt, Alex S; Panzarino, Alexandra M et al. (2018) Deep experimental profiling of microRNA diversity, deployment, and evolution across the Drosophila genus. Genome Res 28:52-65
Lin, Ching-Jung; Hu, Fuqu; Dubruille, Raphaelle et al. (2018) The hpRNA/RNAi Pathway Is Essential to Resolve Intragenomic Conflict in the Drosophila Male Germline. Dev Cell 46:316-326.e5
Jee, David; Yang, Jr-Shiuan; Park, Sun-Mi et al. (2018) Dual Strategies for Argonaute2-Mediated Biogenesis of Erythroid miRNAs Underlie Conserved Requirements for Slicing in Mammals. Mol Cell 69:265-278.e6
Kan, Lijuan; Grozhik, Anya V; Vedanayagam, Jeffrey et al. (2017) The m6A pathway facilitates sex determination in Drosophila. Nat Commun 8:15737
Kondo, Shu; Vedanayagam, Jeffrey; Mohammed, Jaaved et al. (2017) New genes often acquire male-specific functions but rarely become essential in Drosophila. Genes Dev 31:1841-1846
Lin, Ching-Jung; Wen, Jiayu; Bejarano, Fernando et al. (2017) Characterization of a TUTase/RNase complex required for Drosophila gametogenesis. RNA 23:284-296

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