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Cloning and expression of the cDNA encoding rat caspase-2.

Authors: Sato, N  Milligan, CE  Uchiyama, Y  Oppenheim, RW 
Citation: Sato N, etal., Gene 1997 Nov 20;202(1-2):127-32.
Pubmed: (View Article at PubMed) PMID:9427555

We have isolated cDNA clones for rat caspase-2 (also called Nedd2/Ich-1), that encodes a protein similar to interleukin-1beta-converting enzyme (ICE) and the product of the nematode Caenorhabditis elegans cell death gene ced-3 both of which play an important role in programmed cell death (PCD). The rat caspase-2 cDNA clones have an open reading frame (ORF) of 452 amino acids (aa). The predicted aa sequence of rat caspase-2 is highly similar to that of mouse Nedd2 (97.3%) and human Ich-1L (91.3%). The aa sequence QACRG containing the active Cys residue, that is necessary for the proteolytic activity of ICE/Ced-3 (caspase) family of proteases, is also conserved in rat caspase-2. Rat caspase-2 also has several Asp residues in the amino and carboxyl cleavage regions similar to other caspase family proteins. We have developed PC12 cells carrying an on/off switching cassette of caspase-2 (named PC-Nd cells), which contains the neo gene flanked by a pair of loxP sites, the Cre-specific recognition sequence of 34 nucleotides (nt), that lies between the promoter and the caspase-2 cDNA. This expression cassette was designed to express the neo gene initially and to turn on the expression of caspase-2 by site-specific recombinase Cre-mediated excisional deletion of the neo gene. After infection with Cre-producing recombinant adenovirus (re-Ad), the expression of caspase-2 was highly induced in PC-Nd cells and presumptive actively processed fragments of caspase-2 were also observed. This gene activation strategy of caspase-2 will be useful for the study of the biological effects of caspase family proteins in PCD.

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CRRD Object Information
CRRD ID: 68881
Created: 2001-10-10
Species: All species
Last Modified: 2001-10-10
Status: ACTIVE



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