SAFB2 Enables the Processing of Suboptimal Stem-Loop Structures in Clustered Primary miRNA Transcripts

Mol Cell. 2020 Jun 4;78(5):876-889.e6. doi: 10.1016/j.molcel.2020.05.011.

Abstract

Many microRNAs (miRNAs) are generated from primary transcripts containing multiple clustered stem-loop structures that are thought to be recognized and cleaved by the Microprocessor complex as independent units. Here, we uncover an unexpected mode of processing of the bicistronic miR-15a-16-1 cluster. We find that the primary miR-15a stem-loop is not processed on its own but that the presence of the neighboring primary miR-16-1 stem-loop on the same transcript can compensate for this deficiency in cis. Using a CRISPR/Cas9 screen, we identify SAFB2 (scaffold attachment factor B2) as an essential co-factor in this miR-16-1-assisted pri-miR-15 cleavage and describe SAFB2 as an accessory protein of the Microprocessor. Notably, SAFB2-mediated cleavage expands to other clustered pri-miRNAs, indicating a general mechanism. Together, our study reveals an unrecognized function of SAFB2 in miRNA processing and suggests a scenario in which SAFB2 enables the binding and processing of suboptimal Microprocessor substrates in clustered primary miRNA transcripts.

Keywords: DGCR8; DROSHA; ERH; Microprocessor; SAFB; cluster assistance; miRNA; microRNA; microRNA biogenesis; pri-miRNA processing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Nucleus / metabolism
  • HEK293 Cells
  • Humans
  • Inverted Repeat Sequences / genetics
  • Inverted Repeat Sequences / physiology
  • Matrix Attachment Region Binding Proteins / genetics
  • Matrix Attachment Region Binding Proteins / metabolism*
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Nuclear Matrix-Associated Proteins / genetics
  • Nuclear Matrix-Associated Proteins / metabolism*
  • Nucleic Acid Conformation
  • RNA Processing, Post-Transcriptional / genetics
  • RNA-Binding Proteins / metabolism
  • Receptors, Estrogen / genetics
  • Receptors, Estrogen / metabolism*

Substances

  • MIRN15 microRNA, human
  • MIRN16 microRNA, human
  • Matrix Attachment Region Binding Proteins
  • MicroRNAs
  • Nuclear Matrix-Associated Proteins
  • RNA-Binding Proteins
  • Receptors, Estrogen
  • SAFB2 protein, human