Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II | Zendy

Jesse R. Zamudio | Zendy; Timothy Kelly | Zendy; Phillip A. Sharp | Zendy

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Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II

Author(s) -

Jesse R. Zamudio,

Timothy Kelly,

Phillip A. Sharp

Publication year - 2014

Publication title -

cell

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 26.304

H-Index - 776

eISSN - 1097-4172

pISSN - 0092-8674

DOI - 10.1016/j.cell.2014.01.041

Subject(s) - biology , argonaute , polymerase , rna polymerase ii , rna , genetics , microbiology and biotechnology , rna interference , promoter , gene , gene expression

Argonaute (Ago) proteins mediate posttranscriptional gene repression by binding guide miRNAs to regulate targeted RNAs. To confidently assess Ago-bound small RNAs, we adapted a mouse embryonic stem cell system to express a single epitope-tagged Ago protein family member in an inducible manner. Here, we report the small RNA profile of Ago-deficient cells and show that Ago-dependent stability is a common feature of mammalian miRNAs. Using this criteria and immunopurification, we identified an Ago-dependent class of noncanonical miRNAs derived from protein-coding gene promoters, which we name transcriptional start site miRNAs (TSS-miRNAs). A subset of promoter-proximal RNA polymerase II (RNAPII) complexes produces hairpin RNAs that are processed in a DiGeorge syndrome critical region gene 8 (Dgcr8)/Drosha-independent but Dicer-dependent manner. TSS-miRNA activity is detectable from endogenous levels and following overexpression of mRNA constructs. Finally, we present evidence of differential expression and conservation in humans, suggesting important roles in gene regulation.

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