Tissue-selective effects of nucleolar stress and rDNA damage in developmental disorders

Nature. 2018 Feb 1;554(7690):112-117. doi: 10.1038/nature25449. Epub 2018 Jan 24.

Abstract

Many craniofacial disorders are caused by heterozygous mutations in general regulators of housekeeping cellular functions such as transcription or ribosome biogenesis. Although it is understood that many of these malformations are a consequence of defects in cranial neural crest cells, a cell type that gives rise to most of the facial structures during embryogenesis, the mechanism underlying cell-type selectivity of these defects remains largely unknown. By exploring molecular functions of DDX21, a DEAD-box RNA helicase involved in control of both RNA polymerase (Pol) I- and II-dependent transcriptional arms of ribosome biogenesis, we uncovered a previously unappreciated mechanism linking nucleolar dysfunction, ribosomal DNA (rDNA) damage, and craniofacial malformations. Here we demonstrate that genetic perturbations associated with Treacher Collins syndrome, a craniofacial disorder caused by heterozygous mutations in components of the Pol I transcriptional machinery or its cofactor TCOF1 (ref. 1), lead to relocalization of DDX21 from the nucleolus to the nucleoplasm, its loss from the chromatin targets, as well as inhibition of rRNA processing and downregulation of ribosomal protein gene transcription. These effects are cell-type-selective, cell-autonomous, and involve activation of p53 tumour-suppressor protein. We further show that cranial neural crest cells are sensitized to p53-mediated apoptosis, but blocking DDX21 loss from the nucleolus and chromatin rescues both the susceptibility to apoptosis and the craniofacial phenotypes associated with Treacher Collins syndrome. This mechanism is not restricted to cranial neural crest cells, as blood formation is also hypersensitive to loss of DDX21 functions. Accordingly, ribosomal gene perturbations associated with Diamond-Blackfan anaemia disrupt DDX21 localization. At the molecular level, we demonstrate that impaired rRNA synthesis elicits a DNA damage response, and that rDNA damage results in tissue-selective and dosage-dependent effects on craniofacial development. Taken together, our findings illustrate how disruption in general regulators that compromise nucleolar homeostasis can result in tissue-selective malformations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Benzothiazoles / pharmacology
  • Cell Nucleolus / drug effects
  • Cell Nucleolus / genetics
  • Cell Nucleolus / metabolism*
  • Cell Nucleolus / pathology*
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Cell Nucleus / pathology
  • Chromatin / metabolism
  • DEAD-box RNA Helicases / deficiency
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism
  • DNA Damage*
  • DNA, Ribosomal / genetics
  • DNA, Ribosomal / metabolism*
  • DNA-Directed RNA Polymerases / deficiency
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • HeLa Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Mandibulofacial Dysostosis / embryology
  • Mandibulofacial Dysostosis / genetics*
  • Mandibulofacial Dysostosis / pathology*
  • Mice
  • Naphthyridines / pharmacology
  • Neural Crest / enzymology
  • Neural Crest / pathology
  • Nuclear Proteins / deficiency
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Organ Specificity
  • Phenotype
  • Phosphoproteins / deficiency
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Protein Transport / drug effects
  • RNA Helicases / metabolism
  • RNA Polymerase I / antagonists & inhibitors
  • RNA, Ribosomal / biosynthesis
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism
  • Ribosomal Proteins / biosynthesis
  • Ribosomal Proteins / genetics
  • Ribosomes / genetics
  • Ribosomes / metabolism
  • Skull / pathology
  • Stress, Physiological* / drug effects
  • Tumor Suppressor Protein p53 / metabolism
  • Xenopus
  • Zebrafish / embryology
  • Zebrafish Proteins / deficiency

Substances

  • Benzothiazoles
  • CX 5461
  • Chromatin
  • DNA, Ribosomal
  • Intracellular Signaling Peptides and Proteins
  • Naphthyridines
  • Nuclear Proteins
  • Phosphoproteins
  • RNA, Ribosomal
  • Ribosomal Proteins
  • TCOF1 protein, Xenopus
  • TCOF1 protein, human
  • Tcof1 protein, mouse
  • Tumor Suppressor Protein p53
  • Zebrafish Proteins
  • DNA-Directed RNA Polymerases
  • POLR1D protein, human
  • RNA Polymerase I
  • DDX21 protein, human
  • DEAD-box RNA Helicases
  • RNA Helicases
  • ddx21 protein, zebrafish