title: CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45.
abstract: Induction of apoptosis by death receptors such as Fas or tumour necrosis factor (TNF) R1 leads to distinct changes in cell morphology, activation of the caspase protease cascade, and the degradation of nuclear chromatin by activated nucleases. Here, we describe the purification and cDNA cloning of a novel 40 kDa endonuclease from Jurkat cells that is activated by caspases. This protein, designated caspase-activated nuclease (CPAN), is sufficient to degrade naked DNA and to induce apoptotic morphology and DNA fragmentation in naive nuclei. CPAN is highly homologous to a recently described mouse nuclease, CAD [1], and may represent the human homologue. Our data on the human cDNA as well as additional data on the mouse homologue suggest that a 30 amino-acid portion of the recently published mouse sequence [1] is incorrect. We show that the activity of human CPAN is regulated by DFF45 [2], an inhibitor necessary for CPAN expression and stabilization in an inactive state in living cells. Proteolytic cleavage of DFF45 by caspases in vitro leads to dissociation of DFF45 fragments from CPAN and activation of CPAN as an endonuclease. CPAN is a tightly regulated endonuclease with unique characteristics that might represent a distinctive family of endonucleases.
authors.affiliation: Chiron Corporation, 4560 Horton Street, Emeryville, California 94608, USA. rob_halenbeck@cc.chiron.com
authors.forename: R
authors.initials: R
authors.lastname: Halenbeck
authors.forename: H
authors.initials: H
authors.lastname: MacDonald
authors.forename: A
authors.initials: A
authors.lastname: Roulston
authors.forename: T T
authors.initials: TT
authors.lastname: Chen
authors.forename: L
authors.initials: L
authors.lastname: Conroy
authors.forename: L T
authors.initials: LT
authors.lastname: Williams
mesh_terms: D000595:Amino Acid Sequence; D051017:Apoptosis Regulatory Proteins; D053148:Caspase 3; D020169:Caspases; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D018076:DNA, Complementary; D003851:Deoxyribonucleases / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D011506:Proteins / Q000502:physiology*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
chemical_list: D051017:Apoptosis Regulatory Proteins; D018076:DNA, Complementary; D011506:Proteins; C109664:caspase-activated DNase inhibitor; D003851:Deoxyribonucleases; D004720:Endonucleases; C109663:caspase-activated deoxyribonuclease; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases