An Exportin-1-dependent microRNA biogenesis pathway during human cell quiescence

Proc Natl Acad Sci U S A. 2017 Jun 20;114(25):E4961-E4970. doi: 10.1073/pnas.1618732114. Epub 2017 Jun 5.

Abstract

The reversible state of proliferative arrest known as "cellular quiescence" plays an important role in tissue homeostasis and stem cell biology. By analyzing the expression of miRNAs and miRNA-processing factors during quiescence in primary human fibroblasts, we identified a group of miRNAs that are induced during quiescence despite markedly reduced expression of Exportin-5, a protein required for canonical miRNA biogenesis. The biogenesis of these quiescence-induced miRNAs is independent of Exportin-5 and depends instead on Exportin-1. Moreover, these quiescence-induced primary miRNAs (pri-miRNAs) are modified with a 2,2,7-trimethylguanosine (TMG)-cap, which is known to bind Exportin-1, and knockdown of Exportin-1 or trimethylguanosine synthase 1, responsible for (TMG)-capping, inhibits their biogenesis. Surprisingly, in quiescent cells Exportin-1-dependent pri-miR-34a is present in the cytoplasm together with a small isoform of Drosha, implying the existence of a different miRNA processing pathway in these cells. Our findings suggest that during quiescence the canonical miRNA biogenesis pathway is down-regulated and specific miRNAs are generated by an alternative pathway to regulate genes involved in cellular growth arrest.

Keywords: (TMG)-cap; XPO1; XPO5; pri-miRNA; quiescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosynthetic Pathways / drug effects
  • Biosynthetic Pathways / genetics*
  • Cell Line
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Proliferation / genetics*
  • Cytoplasm / genetics
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Exportin 1 Protein
  • Guanosine / analogs & derivatives
  • Guanosine / pharmacology
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Karyopherins / genetics*
  • MicroRNAs / genetics*
  • Receptors, Cytoplasmic and Nuclear / genetics*

Substances

  • Karyopherins
  • MicroRNAs
  • Receptors, Cytoplasmic and Nuclear
  • Guanosine
  • N(2),N(2),7-trimethylguanosine