Genetic interactions between Brca1 and Gadd45a in centrosome duplication, genetic stability, and neural tube closure

J Biol Chem. 2004 Jul 9;279(28):29606-14. doi: 10.1074/jbc.M312279200. Epub 2004 May 3.

Abstract

GADD45a is a transcription target of the breast tumor suppressor gene BRCA. It was recently shown that mouse embryonic fibroblast cells carrying a targeted deletion of exon 11 of Brca1 (Brca1(Delta11/Delta11)) or a Gadd45A-null mutation (Gadd45a(-/-)) suffer centrosome amplification. To study genetic interactions between these genes during centrosome duplication, we generated Brca1(Delta11/Delta)(11)Gadd45a(-/-) mice by crossing each mutant. We found that all Brca1(Delta11/Delta11)Gadd45a(-/-) embryos at embryonic days 9.5-10.5 were exencephalic and exhibited a high incidence of apoptosis accompanied by altered levels of BAX, BCL-2, and p53. The trigger for these events is likely the genetic instability arising from centrosome amplification that is associated, at least in part, with decreased expression of the NIMA-related kinase NEK2. We demonstrate that small interfering RNA-mediated suppression of Brca1 decreased Nek2 more dramatically in Gadd45a(-/-) cells than in wild-type cells and, conversely, that overexpression of Brca1 and/or Gadd45a up-regulated transcription of Nek2. Furthermore, we show that overexpression of Nek2 in Brca1-specific small interfering RNA-treated wild-type and Gadd45a(-/-) cells repressed abnormal centrosome amplification. These observations suggest that NEK2 plays a role in mediating the actions of BRCA1 and GADD45a in regulating centrosome duplication and in maintaining genetic stability.

MeSH terms

  • Animals
  • Apoptosis / physiology
  • BRCA1 Protein / genetics
  • BRCA1 Protein / metabolism*
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Cells, Cultured
  • Central Nervous System / anatomy & histology*
  • Central Nervous System / embryology*
  • Centrosome / metabolism*
  • Embryo, Mammalian / anatomy & histology
  • Embryo, Mammalian / physiology
  • Female
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Gene Expression Regulation, Developmental
  • Genomic Instability*
  • Gestational Age
  • Humans
  • Mice
  • Mice, Knockout
  • NIMA-Related Kinases
  • Neural Tube Defects / genetics
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Pregnancy
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • RNA, Small Interfering / metabolism
  • Transcription, Genetic
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • BRCA1 Protein
  • Cell Cycle Proteins
  • GADD45A protein, human
  • Gadd45a protein, mouse
  • Nuclear Proteins
  • RNA, Small Interfering
  • Tumor Suppressor Protein p53
  • NEK2 protein, human
  • NIMA-Related Kinases
  • Nek2 protein, mouse
  • Protein Serine-Threonine Kinases