Nucleases in programmed cell death

Methods Enzymol. 2008;442:271-87. doi: 10.1016/S0076-6879(08)01414-6.

Abstract

DNA degradation is one of the hallmarks of programmed cell death, or apoptosis. Recent analyses of this process revealed that apoptotic DNA degradation is mediated by two independent mechanisms. First, the caspase-activated DNase (CAD) cell autonomously cleaves DNA into nucleosomal units in dying cells. Then, after the apoptotic cells are engulfed by macrophages, the fragmented DNA is further degraded by DNase II in the lysosomes of the macrophages. This chapter describes assay procedures for CAD and DNase II. It includes biochemical methods for quantifying DNase activity and cell culture systems to follow cell-autonomous and noncell-autonomous DNA degradation. These techniques are useful for studying DNases that are involved in programmed cell death and for following the engulfment of apoptotic cells by phagocytes.

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Apoptosis / physiology*
  • Cell Line
  • Cells, Cultured
  • DNA Fragmentation
  • Deoxyribonucleases / metabolism*
  • Endodeoxyribonucleases / metabolism*
  • HeLa Cells
  • Humans
  • Lymphocytes / cytology
  • Lymphocytes / metabolism

Substances

  • Deoxyribonucleases
  • Endodeoxyribonucleases
  • caspase-activated deoxyribonuclease
  • deoxyribonuclease II