Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

• Published in: Journal of cellular biochemistry 1999-01, Vol.72 (1), p.35-46
• Main Authors: Martelli, Alberto M., Bortul, Roberta, Fackelmayer, Frank O., Tazzari, Pier Luigi, Bareggi, Renato, Narducci, Paola, Zweyer, Marina
• Format: Article
• Language: English
• Subjects: apoptosis; Apoptosis - drug effects; Blotting, Western; Camptothecin - pharmacology; Cell Nucleus - metabolism; Cell Nucleus - ultrastructure; Electrophoresis, Gel, Two-Dimensional; Enzyme Inhibitors - pharmacology; Fluorescent Antibody Technique; HL-60 Cells; Humans; immunocytochemistry; Microscopy, Electron; Neoplasm Proteins - analysis; nuclear matrix; Nuclear Matrix - chemistry; Nuclear Proteins - analysis; protein degradation; scaffold/matrix-associated regions; Topoisomerase I Inhibitors
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/(SICI)1097-4644(19990101)72:1<35::AID-JCB5>3.0.CO;2-S

We have characterized the nuclear matrix‐intermediate filament fraction from control and apoptotic HL‐60 cells. Apoptosis was induced by exposure to the topoisomerase I inhibitor, camptothecin. By means of two‐dimensional polyacrylamide gel electrophoresis, striking qualitative and quantitative differences were seen in the protein composition of the nuclear matrix–intermediate filament fraction obtained from apoptotic cells in comparison with controls. Western blotting analysis of apoptotic nuclear matrix proteins revealed degradation of some (topoisomerase IIα, SAF‐A) but not other (SATB1 and nucleolin) components. Moreover, immunofluorescent staining for typical matrix antigens (NuMA protein, lamin B, SC‐35) showed that in 35–40% of the structures prepared from apoptotic samples, marked changes in the subnuclear distribution of these proteins were present. Striking morphological differences between control and apoptotic samples were also detected at the ultrastructural level. These results demonstrate that both biochemical and morphological changes can be detected in the nuclear matrix prepared from apoptotic HL‐60 cells. J. Cell. Biochem. 72:35–46, 1999. © 1999 Wiley‐Liss, Inc.