The major apoptotic endonuclease DFF40/CAD is a deoxyribose-specific and double-strand-specific enzyme |
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Authors: | Jakub Hanus Magdalena Kalinowska-Herok Piotr Widlak |
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Institution: | (1) Department of Experimental and Clinical Radiobiology, Maria Sklodowska-Curie Cancer Center and Institute of Oncology, Branch in Gliwice, 15 Wybrzeże AK, 44-100 Gliwice, Poland;(2) Faculty of Biotechnology, University of Rzeszów, 36-100 Kolbuszowa, Poland |
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Abstract: | DFF40/CAD endonuclease is primarily responsible for internucleosomal DNA cleavage during the terminal stages of apoptosis.
The nuclease specifically introduces DNA double strand breaks into chromatin substrates. Here we performed a detailed study
on the specificity of the nuclease using synthetic single-stranded and double-stranded ribo- and deoxyribo-oligonucleotides
as substrates. We have found that neither single-stranded DNA, single-stranded RNA, double-stranded RNA nor RNA–DNA heteroduplexes
are cleaved by the DFF40/CAD nuclease. Noteworthy, all types of oligonucleotides that are not cleaved by the nuclease inhibit
cleavage of double-stranded DNA. We have also observed that in cells undergoing apoptosis in vivo neither the activation of
DFF40/CAD nor oligonucleosomal chromatin fragmentation was temporally correlated with either total cellular or nuclear RNA
degradation. We conclude that DFF40/CAD is exclusively specific for double-stranded DNA.
Jakub Hanus and Magdalena Kalinowska-Herok contributed equally to the work. |
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Keywords: | Apoptotic nuclease DFF CAD DNase RNase |
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