Human caspase 12 has acquired deleterious mutations

https://doi.org/10.1016/S0006-291X(02)00289-9Get rights and content

Abstract

Caspase 12 has been cloned from rodent cells, in which it mediated apoptosis in response to endoplasmic reticulum stress. Based on experiments with murine cells it was suggested that this caspase plays a central role in the pathogenesis of Alzheimer's disease. By alignment of the murine caspase 12 cDNA with the human genome sequence we localized the human caspase 12 gene at a single locus within the caspase 1/ICE gene cluster on chromosome 11q22.3. RT-PCR and molecular cloning revealed that nine alternatively spliced transcripts of this gene are expressed. A frame shift mutation and a premature stop codon which is present in all splice variants preclude the expression of a full length protein. An additional loss-of-function mutation within the SHG box, a critical site in caspases, prohibits any proteins, if they are produced, from acting catalytically. Based on our data we conclude that functional caspase 12 is lost in humans and that it can therefore not play a role in Alzheimer's disease.

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Materials and methods

cDNA synthesis and polymerase chain reaction (PCR). Poly(A)+ RNA (0.5 μg) from multiple tissues (Clontech, USA) and 1.5 μg oligo(dT)15 primer (0.5 μg/μl) were denatured for 15 min at 70 °C in 11 μl diethyl pyrocarbonate-treated water (ADDEPC). For cDNA synthesis the following reagents were added: 5 μl 5× AMV buffer (Roche, Switzerland), 2.5 μl dNTP mix 10 mM each (TaKaRa, Japan), 1.5 μl RNasin (40 U/μl) (Roche, Switzerland), 1.5 μl AMV reverse transcriptase (10 U/μl) (Roche, Switzerland), and 2 μl ADDEPC and

Results and discussion

BLAST search of the human genome working draft [12] (http://www.ncbi.nlm.nih.gov/genome/seq/HsBlast.html), which covers 98.5% of the human genome sequence, with the full length sequence of the murine caspase 12 cDNA (GenBank Acc No. NM_009808) [6], revealed a single caspase 12 locus consisting of at least eight putative exons on the chromosome 11q22.3. The gene of caspase 12 is located within a region of genes of the interleukin-1β converting enzyme (ICE) subfamily of caspases (caspases 1, 4,

Acknowledgements

We thank Heidi Rossiter for a critical reading of the manuscript and Aumaid Uthman for the technical advice.

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These authors contribute equally to this paper.

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