Chromatin structure and endonuclease sensitivity in human leukemic cell lines.
The relationship between chromatin structure and endonuclease sensitivity was investigated. The cells used in this study were a) human myelogenous leukemic cell lines (HL-60, ML-I, U-937, THP-I) (Group I), which produced internucleosomal DNA cleavage, and b) human T-cell leukemia (MOLT-4), erythroleukemia (K562), glioblastoma (T98G, U87MG) and glioma (KG-1-C) cell lines (Group II), which produced no internucleosomal DNA cleavage, upon treatment with various apoptosis-inducing agents. When the nuclei, isolated from these cells were digested with micrococcal nuclease, chromatin DNA was cleaved into oligonucleosomal units. Although sensitivity to micrococcal nuclease considerably differed from cell to cell, Group I cells were generally more sensitive to micrococcal nuclease digestion than Group II cells. Similar sensitivity to DNase I was observed in both groups of cells. Acid-urea polyacrylamide gel electrophoresis of histone fractions from control and apoptosing HL-60 cells (induced either by hydrogen peroxide or UV irradiation) revealed no significant change in the relative composition of five major histones, indicating the absence of selective degradation of histone HI, but rather the nonspecific degradation of many nuclear proteins. These data suggest a difference in a chromatin structure between Group I and II cells, which might result in the selective production of internucleosomal DNA cleavage only in Group I cells.
Kuribayashi
N
N
First Department of Biochemistry, School of Medicine, Showa University, Tokyo, Japan.
Sakagami
H
H
Iida
M
M
Takeda
M
M
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002843:Chromatin / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism* / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D004789:Enzyme Activation; D018922:HL-60 Cells / Q000201:enzymology / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D008836:Micrococcal Nuclease / Q000378:metabolism* / Q000494:pharmacology; D009369:Neoplasms / Q000201:enzymology / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured / Q000187:drug effects
D002843:Chromatin; D006657:Histones; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease; D010450:Endopeptidases