TY - JOUR
T1 - Early necrotic DNA degradation
T2 - Presence of blunt-ended DNA breaks, 3′ and 5′ overhangs in apoptosis, but only 5′ overhangs in early necrosis
AU - Didenko, Vladimir V.
AU - Ngo, Hop
AU - Baskin, David S.
N1 - Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2003/5/1
Y1 - 2003/5/1
N2 - The structure of DNA breaks in early necrosis was analyzed and compared with apoptotic DNA degradation using in vivo and cell culture models. Early necrosis (1 hour after cell death) was produced in vivo by the freezing-thawing of rat thymus and in cell culture of Jurkat cells. Apoptosis was induced in the same cell types using dexamethasone for thymus and staurosporine for Jurkat cells. Selective detection of double-strand DNA breaks with blunt ends was performed by in situ ligation. Blunt-ended breaks bearing 5′ phosphates were detected in apoptotic but not in early necrotic cells. Pretreatment of apoptotic and necrotic tissue with Klenow enzyme with or without added dNTPs reduced all 3′ or 5′ overhangs to blunt ends. Subsequent in situ ligation with blunt-ended probes revealed no 3′ overhangs in necrotic cells. However double-strand cuts with 5′ overhangs were abundant in necrotic DNA. 5′ Overhangs were also detected in apoptotic cells. Presence of exclusively 5′ overhangs in early necrosis with absence of a variety of possible DNA ends, suggests the existence of a specific orderly mechanism of DNA degradation.
AB - The structure of DNA breaks in early necrosis was analyzed and compared with apoptotic DNA degradation using in vivo and cell culture models. Early necrosis (1 hour after cell death) was produced in vivo by the freezing-thawing of rat thymus and in cell culture of Jurkat cells. Apoptosis was induced in the same cell types using dexamethasone for thymus and staurosporine for Jurkat cells. Selective detection of double-strand DNA breaks with blunt ends was performed by in situ ligation. Blunt-ended breaks bearing 5′ phosphates were detected in apoptotic but not in early necrotic cells. Pretreatment of apoptotic and necrotic tissue with Klenow enzyme with or without added dNTPs reduced all 3′ or 5′ overhangs to blunt ends. Subsequent in situ ligation with blunt-ended probes revealed no 3′ overhangs in necrotic cells. However double-strand cuts with 5′ overhangs were abundant in necrotic DNA. 5′ Overhangs were also detected in apoptotic cells. Presence of exclusively 5′ overhangs in early necrosis with absence of a variety of possible DNA ends, suggests the existence of a specific orderly mechanism of DNA degradation.
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U2 - 10.1016/S0002-9440(10)64291-5
DO - 10.1016/S0002-9440(10)64291-5
M3 - Article
C2 - 12707041
AN - SCOPUS:0037407775
VL - 162
SP - 1571
EP - 1578
JO - American Journal of Pathology
JF - American Journal of Pathology
SN - 0002-9440
IS - 5
ER -