Lack of WDR36 leads to preimplantation embryonic lethality in mice and delays the formation of small subunit ribosomal RNA in human cells in vitro

Hum Mol Genet. 2011 Feb 1;20(3):422-35. doi: 10.1093/hmg/ddq478. Epub 2010 Nov 3.

Abstract

Mutations in WD repeat domain 36 gene (WDR36) play a causative role in some forms of primary open-angle glaucoma, a leading cause of blindness worldwide. WDR36 is characterized by the presence of multiple WD40 repeats and shows homology to Utp21, an essential protein component of the yeast small subunit (SSU) processome required for maturation of 18S rRNA. To clarify the functional role of WDR36 in the mammalian organism, we generated and investigated mutant mice with a targeted deletion of Wdr36. In parallel experiments, we used RNA interference to deplete WDR36 mRNA in mouse embryos and cultured human trabecular meshwork (HTM-N) cells. Deletion of Wdr36 in the mouse caused preimplantation embryonic lethality, and essentially similar effects were observed when WDR36 mRNA was depleted in mouse embryos by RNA interference. Depletion of WDR36 mRNA in HTM-N cells caused apoptotic cell death and upregulation of mRNA for BAX, TP53 and CDKN1A. By immunocytochemistry, staining for WDR36 was observed in the nucleolus of cells, which co-localized with that of nucleolar proteins such as nucleophosmin and PWP2. In addition, recombinant and epitope-tagged WDR36 localized to the nucleolus of HTM-N cells. By northern blot analysis, a substantial decrease in 21S rRNA, the precursor of 18S rRNA, was observed following knockdown of WDR36. In addition, metabolic-labeling experiments consistently showed a delay of 18S rRNA maturation in WDR36-depleted cells. Our results provide evidence that WDR36 is an essential protein in mammalian cells which is involved in the nucleolar processing of SSU 18S rRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Blastocyst / physiology*
  • Blotting, Northern
  • Cell Nucleolus / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / genetics
  • Embryo Loss / genetics*
  • Embryonic Development / genetics*
  • Eye Proteins / genetics*
  • Eye Proteins / physiology*
  • Fluorescent Antibody Technique
  • Genes, p53
  • Glaucoma, Open-Angle / genetics
  • Humans
  • Mice
  • Mice, Knockout
  • Mutation
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • RNA Interference
  • RNA, Ribosomal / metabolism
  • RNA, Ribosomal, 18S / metabolism*
  • bcl-2-Associated X Protein / genetics

Substances

  • 21S rRNA
  • Cyclin-Dependent Kinase Inhibitor p21
  • Eye Proteins
  • Nuclear Proteins
  • RNA, Ribosomal
  • RNA, Ribosomal, 18S
  • WDR36 protein, mouse
  • bcl-2-Associated X Protein