Distinct RNA polymerase transcripts direct the assembly of phase-separated DBC1 nuclear bodies in different cell lines

Mol Biol Cell. 2021 Nov 1;32(21):ar33. doi: 10.1091/mbc.E21-02-0081. Epub 2021 Sep 8.

Abstract

The mammalian cell nucleus is a highly organized organelle that contains membrane-less structures referred to as nuclear bodies (NBs). Some NBs carry specific RNA types that play architectural roles in their formation. Here, we show two types of RNase-sensitive DBC1-containing NBs, DBC1 nuclear body (DNB) in HCT116 cells and Sam68 nuclear body (SNB) in HeLa cells, that exhibit phase-separated features and are constructed using RNA polymerase I or II transcripts in a cell type-specific manner. We identified additional protein components present in DNB by immunoprecipitation-mass spectrometry, some of which (DBC1 and heterogeneous nuclear ribonucleoprotein L [HNRNPL]) are required for DNB formation. The rescue experiment using the truncated HNRNPL mutants revealed that two RNA-binding domains and intrinsically disordered regions of HNRNPL play significant roles in DNB formation. All these domains of HNRNPL promote in vitro droplet formation, suggesting the need for multivalent interactions between HNRNPL and RNA as well as proteins in DNB formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Cell Nucleus / metabolism
  • DNA-Binding Proteins / metabolism
  • DNA-Directed RNA Polymerases / metabolism*
  • DNA-Directed RNA Polymerases / physiology
  • HeLa Cells
  • Humans
  • Nuclear Bodies / metabolism*
  • Nuclear Bodies / physiology
  • RNA-Binding Proteins / metabolism
  • Ribonucleases / metabolism
  • Ribonucleoproteins / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • CCAR2 protein, human
  • DNA-Binding Proteins
  • KHDRBS1 protein, human
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • DNA-Directed RNA Polymerases
  • Ribonucleases