Discovery, regulation, and action of the major apoptotic nucleases DFF40/CAD and endonuclease G

Piotr Widlak, William T. Garrard

Research output: Contribution to journalArticle

169 Citations (Scopus)

Abstract

Toward the end of the 20th and beginning of the 21st centuries, clever in vitro biochemical complementation experiments and genetic screens from the laboratories of Xiaodong Warig, Shigekazu Nagata, and Ding Xue led to the discovery of two major apoptotic nucleases, termed DNA fragmentation factor (DFF) or caspase-activated DNase (CAD) and endonuclease C (Endo G). Both endonucleases attack chromatin to yield 3′-hydroxyl groups and 5′-phosphate residues, first at the level of 50-300 kb cleavage products and next at the level of internucleosomal DNA fragmentation, but these nucleases possess completely different cellular locations in normal cells and are regulated in vastly different ways. In non-apoptotic cells, DFF exists in the nucleus as a heterodimer, composed of a 45 kD chaperone and inhibitor subunit (DFF45) [also called inhibitor of CAD (ICAD-L)] and a 40 kD latent nuclease subunit (DFF40/CAD). Apoptotic activation of caspase-3 or -7 results in the cleavage of DFF45/ICAD and release of active DFF40/CAD nuclease. DFF40's nuclease activity is further activated by specific chromosomal proteins, such as histone H1, HMGB1/2, and topoisomerase II. DFF is regulated by multiple pre- and post-activation fail-safe steps, which include the requirements for DFF45/ICAD, Hsp70, and Hsp40 proteins to mediate appropriate folding during translation to generate a potentially activatable nuclease, and the synthesis in stoichiometric excess of the inhibitors (DFF45/35; ICAD-S/L). By contrast, Endo G resides in the mitochondrial intermembrane space in normal cells, and is released into the nucleus upon apoptotic disruption of mitochondrial membrane permeability in association with co-activators such as apoptosis-inducing factor (AIF). Understanding further regulatory check-points involved in safeguarding non-apoptotic cells against accidental activation of these nucleases remain as future challenges, as well as designing ways to selectively activate these nucleases in tumor cells.

Original languageEnglish (US)
Pages (from-to)1078-1087
Number of pages10
JournalJournal of Cellular Biochemistry
Volume94
Issue number6
DOIs
StatePublished - Apr 15 2005

Fingerprint

DNA Fragmentation
Chemical activation
Endonucleases
Cells
DNA
Apoptosis Inducing Factor
HMGB1 Protein
Type II DNA Topoisomerase
Deoxyribonucleases
Caspase 3
Hydroxyl Radical
Histones
Chromatin
Mitochondrial Membranes
Tumors
Proteins
Phosphates
Association reactions
Permeability
Membranes

Keywords

  • Apoptotic chromosome condensation
  • Apoptotic nuclease mechanisms
  • DNA degradation
  • Higher-order chromatin cleavage
  • Nucleosomes
  • Regulation of apoptotic nucleases

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Discovery, regulation, and action of the major apoptotic nucleases DFF40/CAD and endonuclease G. / Widlak, Piotr; Garrard, William T.

In: Journal of Cellular Biochemistry, Vol. 94, No. 6, 15.04.2005, p. 1078-1087.

Research output: Contribution to journalArticle

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