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

• Published in: Journal of proteomics 2012-06, Vol.75 (11), p.3291-3303
• Main Authors: Navakauskienė, Rūta, Treigytė, Gražina, Borutinskaitė, Veronika-Viktorija, Matuzevičius, Dalius, Navakauskas, Dalius, Magnusson, Karl-Eric
• Format: Article
• Language: English
• Subjects: Ageing, cell death; Alpha-DB; Apoptosis; Biological and medical sciences; caspases; cell membranes; Cell physiology; cytoplasm; cytosol; Diverse techniques; Fundamental and applied biological sciences. Psychology; gels; Hematologic and hematopoietic diseases; humans; image analysis; leukemia; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Leukemic cells; mass spectrometry; Medical sciences; MEDICIN; MEDICINE; Molecular and cellular biology; myosin; signal transduction; transport proteins
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2012.03.041

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. [Display omitted] ► In apoptotic cells α-DB localizes between the cell membrane, cytosol and nucleus. ► α-DB associates with structural, transport, ER-Golgi and nuclear proteins. ► An apoptosis-induction in HL-60 cells involves a network of α-DB-associated proteins.