title: Primary microRNA processing is functionally coupled to RNAP II transcription in vitro.
abstract: Previous studies in vivo reported that processing of primary microRNA (pri-miRNA) is coupled to transcription by RNA polymerase II (RNAP II) and can occur co-transcriptionally. Here we have established a robust in vivo system in which pri-miRNA is transcribed by RNAP II and processed to pre-miRNA in HeLa cell nuclear extracts. We show that both the kinetics and efficiency of pri-miRNA processing are dramatically enhanced in this system compared to that of the corresponding naked pri-miRNA. Moreover, this enhancement is general as it occurs with multiple pri-miRNAs. We also show that nascent pri-miRNA is efficiently processed before it is released from the DNA template. Together, our work directly demonstrates that transcription and pri-miRNA processing are functionally coupled and establishes the first in vivo model systems for this functional coupling and for co-transcriptional processing.
authors.affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Ave. Boston MA 02115.
authors.forename: Shanye
authors.initials: S
authors.lastname: Yin
authors.affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Ave. Boston MA 02115.
authors.forename: Yong
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Ave. Boston MA 02115.
authors.forename: Robin
authors.initials: R
authors.lastname: Reed
mesh_terms: D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D014158:Transcription, Genetic
chemical_list: D035683:MicroRNAs; D012322:RNA Precursors; D012319:RNA Polymerase II