Identification and developmental expression of inhibitor of caspase-activated DNase (ICAD) in Drosophila melanogaster.

Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print Cited Medium: Print ISSN: 0021-9258 (Print) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE

Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology

Gene Expression Regulation, Developmental*; Deoxyribonucleases/*metabolism ; Drosophila melanogaster/*physiology ; Proteins/*genetics ; Proteins/*metabolism; Amino Acid Sequence ; Animals ; Apoptosis/drug effects ; Apoptosis Regulatory Proteins ; Caspase 3 ; Caspases/metabolism ; Cell Line ; DNA Fragmentation ; Deoxyribonucleases/antagonists & inhibitors ; Deoxyribonucleases/chemistry ; Drosophila melanogaster/embryology ; Drosophila melanogaster/growth & development ; Expressed Sequence Tags ; Female ; Humans ; Insect Proteins/metabolism ; Insect Proteins/pharmacology ; Larva ; Male ; Mice ; Molecular Sequence Data ; Neurons/cytology ; Neurons/physiology ; Proteins/chemistry ; Proteins/pharmacology ; Pupa ; Recombinant Proteins/metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid

DNA fragmentation, a hallmark of apoptosis, is regulated by a specific nuclease called caspase-activated DNase (CAD) and its inhibitor (ICAD). When cell lysates from Drosophila S2 cells were chemically denatured and the denatured proteins were removed after dialysis, the supernatant inhibited Drosophila CAD (dCAD). To identify the inhibitor, we tested recombinant DREP-1, which was previously identified using the Drosophila EST data base and found it also inhibited dCAD DNase. An antibody against DREP-1 inhibited the ICAD activity in the S2 cell extracts, confirming the identification of DREP-1 as a Drosophila homolog of ICAD (dICAD). The recombinant DREP-1/dICAD was cleaved at a specific site by human caspase 3 as well as by extracts prepared from S2 cells undergoing apoptosis. Biochemical fractionation and immunoprecipitation of dICAD from S2 cell extracts indicated that dICAD is complexed with dCAD in proliferating cells. The expression of the caspase-resistant form of dICAD/DREP-1 in a Drosophila neuronal cell line prevented the apoptotic DNA fragmentation. Northern hybridization and the immunohistochemical analyses revealed that the expression of the dICAD gene is developmentally regulated.

0 (Apoptosis Regulatory Proteins)
0 (Insect Proteins)
0 (Proteins)
0 (Recombinant Proteins)
0 (caspase-activated DNase inhibitor)
EC 3.1.- (Deoxyribonucleases)
EC 3.1.- (caspase-activated deoxyribonuclease)
EC 3.4.22.- (CASP3 protein, human)
EC 3.4.22.- (Casp3 protein, mouse)
EC 3.4.22.- (Caspase 3)
EC 3.4.22.- (Caspases)

Date Created: 20000427 Date Completed: 20000816 Latest Revision: 20210209

20231215

10.1074/jbc.M909611199

10781612