title: Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs.
abstract: miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to study and the in vivo kinetics of this process is not known. Here, we present a method for determining the processing kinetics of pri-miRNAs within intact cells over time, using a pulse-chase approach to label transcribed RNA during 15 min, and follow the processing within a 1-hour window after labeling with bromouridine. We show that pri-miRNAs exhibit different processing kinetics ranging from fast over intermediate to slow processing, and we provide evidence that pri-miRNA processing can occur both cotranscriptionally and post-transcriptionally.
authors.affiliation: Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany.
authors.forename: Annita
authors.initials: A
authors.lastname: Louloupi
authors.affiliation: Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Ntini
authors.affiliation: Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Liz
authors.affiliation: Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany.
authors.forename: Ulf Andersson
authors.initials: UA
authors.lastname: Ørom
mesh_terms: D002460:Cell Line; D002843:Chromatin / Q000235:genetics; D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007700:Kinetics; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D035683:MicroRNAs
keywords: miRNA; miRNA biogenesis; pri-miRNA