Studies of the lamin proteinase reveal multiple parallel biochemical pathways during apoptotic execution

Yuri A. Lazebnik, Atsushi Takahashi, Robert D. Moir, Robert D. Goldman, Guy G. Poirier, Scott H Kaufmann, William C. Earnshaw

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Abstract

Although specific proteinases play a critical role in the active phase of apoptosis, their substrates are largely unknown. We previously identified poly(ADP-ribose) polymerase (PARP) as an apoptosis-associated substrate for proteinase(s) related to interleukin 1β-converting enzyme (ICE). Now we have used a cell-free system to characterize proteinase(s) that cleave the nuclear lamins during apoptosis. Lamin cleavage during apoptosis requires the action of a second ICE-like enzyme, which exhibits kinetics of cleavage and a profile of sensitivity to specific inhibitors that is distinct from the PARP proteinase. Thus, multiple ICE-like enzymes are required for apoptotic events in these cell-free extracts. Inhibition of the lamin proteinase with tosyllysine 'chloromethyl ketone' blocks nuclear apoptosis prior to the packaging of condensed chromatin into apoptotic bodies. Under these conditions, the nuclear DNA is fully cleaved to a nucleosomal ladder. Our studies reveal that the lamin proteinase and the fragmentation nuclease function in independent parallel pathways during the final stages of apoptotic execution. Neither pathway alone is sufficient for completion of nuclear apoptosis. Instead, the various activities cooperate to drive the disassembly of the nucleus.

Original languageEnglish (US)
Pages (from-to)9042-9046
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number20
DOIs
StatePublished - Sep 26 1995

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Apoptosis
Caspase 1
Peptide Hydrolases
Lamins
Poly(ADP-ribose) Polymerases
Tosyllysine Chloromethyl Ketone
Cell-Free System
Product Packaging
Enzymes
Cell Extracts
Chromatin
lamin proteinase
DNA

ASJC Scopus subject areas

  • Genetics
  • General

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Studies of the lamin proteinase reveal multiple parallel biochemical pathways during apoptotic execution. / Lazebnik, Yuri A.; Takahashi, Atsushi; Moir, Robert D.; Goldman, Robert D.; Poirier, Guy G.; Kaufmann, Scott H; Earnshaw, William C.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 20, 26.09.1995, p. 9042-9046.

Research output: Contribution to journalArticle

Lazebnik, Yuri A. ; Takahashi, Atsushi ; Moir, Robert D. ; Goldman, Robert D. ; Poirier, Guy G. ; Kaufmann, Scott H ; Earnshaw, William C. / Studies of the lamin proteinase reveal multiple parallel biochemical pathways during apoptotic execution. In: Proceedings of the National Academy of Sciences of the United States of America. 1995 ; Vol. 92, No. 20. pp. 9042-9046.
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