Silencing of Nuclear Mitotic Apparatus protein (NuMA) accelerates the apoptotic disintegration of the nucleus

Apoptosis. 2010 Aug;15(8):936-45. doi: 10.1007/s10495-010-0506-8.

Abstract

One main feature of apoptosis is the sequential degradation of the nuclear structure, including the fragmentation of chromatin and caspase-mediated cleavage of various nuclear proteins. Among these proteins is the Nuclear Mitotic Apparatus protein (NuMA) which plays a specific role in the organization of the mitotic spindle. The exact function of NuMA in the interphase nucleus is unknown, but a number of reports have suggested that it may play a role in chromatin organization and/or gene expression. Here we show that upon cleavage in apoptotic cells, the N-terminal cleavage fragment of NuMA is solubilized while the C-terminal fragment remains associated with the condensed chromatin. Using pancaspase inhibitor z-VAD-fmk and caspase-3 deficient MCF-7 cells, we further show that the solubilization is dependent on caspase-mediated cleavage of NuMA. Finally, the silencing of NuMA by RNAi accelerated nuclear breakdown in apoptotic MCF-7 cells. These results suggest that NuMA may provide structural support in the interphase nucleus by contributing to the organization of chromatin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Chloromethyl Ketones / metabolism
  • Antigens, Nuclear / genetics
  • Antigens, Nuclear / metabolism*
  • Apoptosis / physiology*
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Caspase Inhibitors
  • Cell Cycle Proteins
  • Cell Line, Tumor
  • Cell Nucleus / metabolism*
  • Chromatin / metabolism
  • Cysteine Proteinase Inhibitors / metabolism
  • Gene Silencing*
  • Humans
  • Nuclear Matrix-Associated Proteins / genetics
  • Nuclear Matrix-Associated Proteins / metabolism*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Amino Acid Chloromethyl Ketones
  • Antigens, Nuclear
  • Caspase Inhibitors
  • Cell Cycle Proteins
  • Chromatin
  • Cysteine Proteinase Inhibitors
  • NUMA1 protein, human
  • Nuclear Matrix-Associated Proteins
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Caspase 3