Alpha-Dystrobrevin and its associated proteins in human promyelocytic leukemia cells induced to apoptosis.

Abstract

Dystrobrevin is a dystrophin-related component of the dystrophin-associated protein complex (DAPC). Using alpha-dystrobrevin as indicator, we aimed to elucidate the interaction network of the DAPC with other proteins during apoptosis of promyelocytic HL-60 cells. The precise role(s) of DBs are not known, but we and others have shown that they play a role in intracellular signal transduction and cellular organization. Apoptosis was induced with etoposide in the absence or presence of Z-VAD to block caspase activity, and we then followed the cellular distribution of α-DB and its association with other proteins, using confocal imaging and cell fractions analyses after immune-precipitation with anti-α-DB and mass spectrometry. Confocal imaging revealed distinct spatial relocalizations of α-DB between the cell membrane, cytosol and nucleus after induction of apoptosis. The expression levels of the identified proteins were evaluated with computer-assisted image analysis of the gels. We thus identified associations with structural and transport proteins (tropomyosin, myosin), membrane (ADAM21, syntrophin), ER-Golgi (TGN51, eIF38) and nuclear (Lamins, ribonucleoprotein C1/C2) proteins. These results suggest that apoptosis-induction in HL-60 cells involves not only classical markers of apoptosis but also a network α-DB-associated proteins at the cell membrane, the cytoplasm and nucleus, affecting key cellular transport processes and cellular structure.

• Authors: Rūta Navakauskienė, Gražina Treigytė, Veronika-Viktorija Borutinskaitė, Dalius Matuzevičius, Dalius Navakauskas, Karl-Eric Magnusson
• Journal: Journal of proteomics (J Proteomics) Vol. 75 Issue 11 Pg. 3291-303 (Jun 18 2012) ISSN: 1876-7737 [Electronic] Netherlands
• PMID: 22507200 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier B.V. All rights reserved.

Chemical References

• Antineoplastic Agents, Phytogenic
• Dystrophin-Associated Proteins
• Neoplasm Proteins
• Oligopeptides
• benzyloxycarbonyl-valyl-alanyl-aspartic acid
• dystrobrevin
• Etoposide

Topics

• Antineoplastic Agents, Phytogenic (pharmacology)
• Apoptosis
• Dystrophin-Associated Proteins (metabolism)
• Etoposide (pharmacology)
• HL-60 Cells
• Humans
• Leukemia, Promyelocytic, Acute (drug therapy, metabolism)
• Neoplasm Proteins (metabolism)
• Oligopeptides (pharmacology)