Journal article
The effect of ICAD-S on the formation and intracellular distribution of a nucleolytically active caspase-activated DNase
Nucleic acids research, Vol.30(14), pp.3045-3051
07/15/2002
Handle:
https://hdl.handle.net/2376/106742
PMCID: PMC135751
PMID: 12136086
Abstract
We show here that co-expression of murine CAD with either ICAD-L or ICAD-S in
Escherichia coli
as well as mammalian cells leads to a functional DFF complex, which after caspase-3 activation releases a nucleolytically active DNase. The chaperone activity of ICAD-S is between one and two orders of magnitude less effective than that of ICAD-L, as deduced from cleavage experiments with different activated recombinant DFF complexes produced in
E.coli
. With nucleolytically active EGFP fusion proteins of CAD it is demonstrated that co-expression of ICAD-S, which lacks the C-terminal domain of ICAD-L, including the NLS, leads to a homogeneous intracellular distribution of the DNase in transfected cells, whereas co-expression of human or murine ICAD-L variants lacking the NLS leads to exclusion of EGFP–CAD from the nuclei in ∼50% of cells. These results attribute a particular importance of the NLS in the long isoform of the inhibitor of CAD for nuclear accumulation of the DFF complex in living cells. It is concluded that ICAD-L and ICAD-S
in vivo
might function as tissue-specific modulators in the regulation of apoptotic DNA degradation by controlling not only the enzymatic activity but also the amount of CAD available in the nuclei of mammalian cells.
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Details
- Title
- The effect of ICAD-S on the formation and intracellular distribution of a nucleolytically active caspase-activated DNase
- Creators
- Sebastian Richard Scholz - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, GermanyChristian Korn - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, GermanyOleg Gimadutdinow - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, GermanyMichael Knoblauch - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, GermanyAlfred Pingoud - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, GermanyGregor Meiss - Institut für Biochemie (FB 08), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, Germany
- Publication Details
- Nucleic acids research, Vol.30(14), pp.3045-3051
- Academic Unit
- Biological Sciences, School of
- Publisher
- Oxford University Press; Oxford, UK
- Identifiers
- 99900546991901842
- Language
- English
- Resource Type
- Journal article