Molecular cloning and characterization of human caspase-activated dnase
Naomi Mukae,Masato Enari,Hideki Sakahira,Yoji Fukuda,Johji Inazawa,Hiroyuki Toh,Shigekazu Nagata +6 more
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TLDR
Overexpression of CAD potentiated DNA fragmentation by apoptotic stimuli in these cell lines, indicating that CAD is responsible for the apoptotic DNA degradation.Abstract:
Caspase-activated DNase (CAD) cleaves chromosomal DNA during apoptosis. Here, we report isolation of two classes of human CAD cDNAs from a human KT-3 leukemic cell cDNA library. One class of cDNA encoded a protein comprising 338 amino acids, which showed a marked similarity to its murine counterpart. In vitro transcription and translation of this cDNA resulted in a functional CAD protein when the protein was synthesized in the presence of its inhibitor (inhibitor of CAD). The other cDNA class contained many deletions, insertions, and point mutations in the sequence corresponding to the coding region, suggesting that it is derived from a pseudogene. The functional CAD gene was localized to human chromosome 1p36.3 by fluorescent in situ hybridization. The CAD mRNA was expressed in a limited number of human tissues, including pancreas, spleen, prostate, and ovary. The expression of the CAD mRNA in human cell lines correlated with their ability to show DNA fragmentation during apoptosis. Overexpression of CAD potentiated DNA fragmentation by apoptotic stimuli in these cell lines, indicating that CAD is responsible for the apoptotic DNA degradation.read more
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Journal ArticleDOI
Apoptotic dna fragmentation
TL;DR: The molecular mechanism of and the physiological roles played by apoptotic DNA fragmentation will be discussed.
Journal ArticleDOI
Initiation of DNA Fragmentation during Apoptosis Induces Phosphorylation of H2AX Histone at Serine 139
TL;DR: It is shown that γ-H2AX formation is an early chromatin modification following initiation of DNA fragmentation during apoptosis, and it is indicated that any apoptotic endonuclease is sufficient to induce this phosphorylated form of H2AX.
Book ChapterDOI
Genetic Control of Programmed Cell Death in the Nematode Caenorhabditis Elegans
TL;DR: The characterization of genes responsible for programmed cell death in C. elegans has defined a molecular genetic pathway that is conserved evolutionarily and provides a basis for understanding programmed cellDeath in more complex organisms, including humans.
Journal ArticleDOI
Degradation of chromosomal DNA during apoptosis
TL;DR: The mechanism and role of DNA degradation during apoptosis is discussed, and several other endonucleases have been suggested as candidate effectors for the apoptotic degradation of chromosomal DNA.
Journal ArticleDOI
Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation
TL;DR: An in vitro system is used to identify a new nuclear factor, designated Acinus, which induces apoptotic chromatin condensation after cleavage by caspase-3 without inducing DNA fragmentation.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction
TL;DR: A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described, providing a pure preparation of undegraded RNA in high yield and can be completed within 4 h.
Book ChapterDOI
Cell death : the significance of apoptosis
TL;DR: It has proved feasible to categorize most if not all dying cells into one or the other of two discrete and distinctive patterns of morphological change, which have, generally, been found to occur under disparate but individually characteristic circumstances.
Journal ArticleDOI
Apoptosis by death factor.
TL;DR: This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, and Culture of Japan and by a Research Grant from the Princess Takamatsu Cancer Research Fund, and performed in part through Special Coordination Funds of the Science and Technology Agency of the Japanese Government.