title: The human PMR1 endonuclease stimulates cell motility by down regulating miR-200 family microRNAs.
abstract: The motility of MCF-7 cells increases following expression of a human PMR1 transgene and the current study sought to identify the molecular basis for this phenotypic change. Ensemble and single cell analyses show increased motility is dependent on the endonuclease activity of hPMR1, and cells expressing active but not inactive hPMR1 invade extracellular matrix. Nanostring profiling identified 14 microRNAs that are downregulated by hPMR1, including all five members of the miR-200 family and others that also regulate invasive growth. miR-200 levels increase following hPMR1 knockdown, and changes in miR-200 family microRNAs were matched by corresponding changes in miR-200 targets and reporter expression. PMR1 preferentially cleaves between UG dinucleotides within a consensus YUGR element when present in the unpaired loop of a stem-loop structure. This motif is present in the apical loop of precursors to most of the downregulated microRNAs, and hPMR1 targeting of pre-miRs was confirmed by their loss following induced expression and increase following hPMR1 knockdown. Introduction of miR-200c into hPMR1-expressing cells reduced motility and miR-200 target gene expression, confirming hPMR1 acts upstream of Dicer processing. These findings identify a new role for hPMR1 in the post-transcriptional regulation of microRNAs in breast cancer cells.
authors.affiliation: Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Shan-Qing
authors.initials: SQ
authors.lastname: Gu
authors.affiliation: Department of Surgery, The Ohio State University, Columbus, OH 43210, USA Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, OH 43210, USA Center for Regenerative Medicine and Cell-Based Therapies, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gallego-Perez
authors.affiliation: Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Sean P
authors.initials: SP
authors.lastname: McClory
authors.affiliation: Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Shi
authors.affiliation: Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Joonhee
authors.initials: J
authors.lastname: Han
authors.affiliation: Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, OH 43210, USA Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
authors.affiliation: Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH 43210, USA schoenberg.3@osu.edu.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Schoenberg
mesh_terms: D002465:Cell Movement / Q000235:genetics*; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D061986:MCF-7 Cells; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D059372:Nucleotide Motifs; D059368:RNA Isoforms / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D059010:Single-Cell Analysis; D019076:Transgenes
chemical_list: C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D059368:RNA Isoforms; D034741:RNA, Small Interfering; D004722:Endoribonucleases; C116181:polysomal RNase 1; C408087:DICER1 protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases