Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis

Nadia Korfali, Sandrine Ruchaud, David Loegering, Delphine Bernard, Colin Dingwall, Scott H Kaufmann, William C. Earnshaw

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Caspases play a key role during apoptotic execution. In an attempt to elucidate the specific role of caspase-7 we generated a chicken DT40 cell line in which both alleles of the gene were disrupted. Viability assays showed that caspase-7-/- clones are more resistant to the common apoptosis-inducing drugs etoposide and staurosporine. Caspase-7-/- cells show a delay in phosphatidylserine externalization and DNA fragmentation as well as cleavage of the caspase substrates poly(ADP-ribose) polymerase 1 and lamins B1 and B2. Caspase affinity labeling and activity assays indicated that deficient cells exhibit a delay in caspase activation compared with wild type DT40 cells, providing an explanation for the differences in apoptotic execution between caspase-7 null and wild type DT40 cells. These results strongly suggest that caspase-7 is involved earlier than other effector caspases in the apoptotic execution process in DT40 B lymphocytes.

Original languageEnglish (US)
Pages (from-to)1030-1039
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number2
DOIs
StatePublished - Jan 9 2004

Fingerprint

Caspase 7
Caspases
Genes
Apoptosis
Enzymes
Assays
Effector Caspases
DNA Cleavage
Staurosporine
Lymphocytes
Poly(ADP-ribose) Polymerases
Phosphatidylserines
DNA Fragmentation
Etoposide
Labeling
Chickens
B-Lymphocytes
Clone Cells
Chemical activation
Alleles

ASJC Scopus subject areas

  • Biochemistry

Cite this

Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis. / Korfali, Nadia; Ruchaud, Sandrine; Loegering, David; Bernard, Delphine; Dingwall, Colin; Kaufmann, Scott H; Earnshaw, William C.

In: Journal of Biological Chemistry, Vol. 279, No. 2, 09.01.2004, p. 1030-1039.

Research output: Contribution to journalArticle

Korfali, Nadia ; Ruchaud, Sandrine ; Loegering, David ; Bernard, Delphine ; Dingwall, Colin ; Kaufmann, Scott H ; Earnshaw, William C. / Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 2. pp. 1030-1039.
@article{4d8326cbf40e4b23b235dd1dc823bb37,
title = "Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis",
abstract = "Caspases play a key role during apoptotic execution. In an attempt to elucidate the specific role of caspase-7 we generated a chicken DT40 cell line in which both alleles of the gene were disrupted. Viability assays showed that caspase-7-/- clones are more resistant to the common apoptosis-inducing drugs etoposide and staurosporine. Caspase-7-/- cells show a delay in phosphatidylserine externalization and DNA fragmentation as well as cleavage of the caspase substrates poly(ADP-ribose) polymerase 1 and lamins B1 and B2. Caspase affinity labeling and activity assays indicated that deficient cells exhibit a delay in caspase activation compared with wild type DT40 cells, providing an explanation for the differences in apoptotic execution between caspase-7 null and wild type DT40 cells. These results strongly suggest that caspase-7 is involved earlier than other effector caspases in the apoptotic execution process in DT40 B lymphocytes.",
author = "Nadia Korfali and Sandrine Ruchaud and David Loegering and Delphine Bernard and Colin Dingwall and Kaufmann, {Scott H} and Earnshaw, {William C.}",
year = "2004",
month = "1",
day = "9",
doi = "10.1074/jbc.M306277200",
language = "English (US)",
volume = "279",
pages = "1030--1039",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",

}

TY - JOUR

T1 - Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis

AU - Korfali, Nadia

AU - Ruchaud, Sandrine

AU - Loegering, David

AU - Bernard, Delphine

AU - Dingwall, Colin

AU - Kaufmann, Scott H

AU - Earnshaw, William C.

PY - 2004/1/9

Y1 - 2004/1/9

N2 - Caspases play a key role during apoptotic execution. In an attempt to elucidate the specific role of caspase-7 we generated a chicken DT40 cell line in which both alleles of the gene were disrupted. Viability assays showed that caspase-7-/- clones are more resistant to the common apoptosis-inducing drugs etoposide and staurosporine. Caspase-7-/- cells show a delay in phosphatidylserine externalization and DNA fragmentation as well as cleavage of the caspase substrates poly(ADP-ribose) polymerase 1 and lamins B1 and B2. Caspase affinity labeling and activity assays indicated that deficient cells exhibit a delay in caspase activation compared with wild type DT40 cells, providing an explanation for the differences in apoptotic execution between caspase-7 null and wild type DT40 cells. These results strongly suggest that caspase-7 is involved earlier than other effector caspases in the apoptotic execution process in DT40 B lymphocytes.

AB - Caspases play a key role during apoptotic execution. In an attempt to elucidate the specific role of caspase-7 we generated a chicken DT40 cell line in which both alleles of the gene were disrupted. Viability assays showed that caspase-7-/- clones are more resistant to the common apoptosis-inducing drugs etoposide and staurosporine. Caspase-7-/- cells show a delay in phosphatidylserine externalization and DNA fragmentation as well as cleavage of the caspase substrates poly(ADP-ribose) polymerase 1 and lamins B1 and B2. Caspase affinity labeling and activity assays indicated that deficient cells exhibit a delay in caspase activation compared with wild type DT40 cells, providing an explanation for the differences in apoptotic execution between caspase-7 null and wild type DT40 cells. These results strongly suggest that caspase-7 is involved earlier than other effector caspases in the apoptotic execution process in DT40 B lymphocytes.

UR - http://www.scopus.com/inward/record.url?scp=0346463061&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0346463061&partnerID=8YFLogxK

U2 - 10.1074/jbc.M306277200

DO - 10.1074/jbc.M306277200

M3 - Article

VL - 279

SP - 1030

EP - 1039

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 2

ER -