title: Granzyme B/perforin-mediated apoptosis of Jurkat cells results in cleavage of poly(ADP-ribose) polymerase to the 89-kDa apoptotic fragment and less abundant 64-kDa fragment.
abstract: Cytotoxic lymphocytes utilize granule associated serine proteases (granzymes) and perforin to induce apoptosis. Although the importance of granzyme B has been established by gene ablation experiments, biochemical events initiated by the granzyme remain enigmatic. We show here that exposure of Jurkat cells to granzyme B and perforin results in cleavage of poly(ADP-ribose) polymerase to an apoptotic 89 kDa fragment and to lesser amounts of a 64 kDa fragment. The 64 kDa fragment is produced directly by granzyme B while the 89 kDa fragment is presumably generated by activated ICE/Ced-3 proteases. Establishing the intracellular function of GrB in the apoptotic response, these results indicate that granzyme B enters perforin treated targets activating the ICE/Ced-3 family proteases which then cleave poly(ADP-ribose) polymerase to its apoptotic fragment. Intracellular granzyme B appears to be translocated to the nucleus where the protease directly cleaves poly(ADP-ribose) polymerase.
authors.affiliation: Department of Medicine, Evanston Hospital, Northwestern University, Illinois 60201, USA. granzyme@merle.acns.nwu.edu
authors.forename: C J
authors.initials: CJ
authors.lastname: Froelich
authors.forename: W L
authors.initials: WL
authors.lastname: Hanna
authors.forename: G G
authors.initials: GG
authors.lastname: Poirier
authors.forename: P J
authors.initials: PJ
authors.lastname: Duriez
authors.forename: D
authors.initials: D
authors.lastname: D'Amours
authors.forename: G S
authors.initials: GS
authors.lastname: Salvesen
authors.forename: E S
authors.initials: ES
authors.lastname: Alnemri
authors.forename: W C
authors.initials: WC
authors.lastname: Earnshaw
authors.forename: G M
authors.initials: GM
authors.lastname: Shah
mesh_terms: D017209:Apoptosis; D002474:Cell-Free System; D053804:Granzymes; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells; D008562:Membrane Glycoproteins / Q000378:metabolism*; D010446:Peptide Fragments / Q000096:biosynthesis*; D054353:Perforin; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D052899:Pore Forming Cytotoxic Proteins; D012697:Serine Endopeptidases / Q000378:metabolism*
chemical_list: D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D011065:Poly(ADP-ribose) Polymerases; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases