title: 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.affiliation: Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania. ruta.navakauskiene@bchi.vu.lt
authors.forename: Rūta
authors.initials: R
authors.lastname: Navakauskienė
authors.forename: Gražina
authors.initials: G
authors.lastname: Treigytė
authors.forename: Veronika-Viktorija
authors.initials: VV
authors.lastname: Borutinskaitė
authors.forename: Dalius
authors.initials: D
authors.lastname: Matuzevičius
authors.forename: Dalius
authors.initials: D
authors.lastname: Navakauskas
authors.forename: Karl-Eric
authors.initials: KE
authors.lastname: Magnusson
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis; D049029:Dystrophin-Associated Proteins / Q000378:metabolism*; D005047:Etoposide / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D049029:Dystrophin-Associated Proteins; D009363:Neoplasm Proteins; D009842:Oligopeptides; C517629:benzyloxycarbonyl-valyl-alanyl-aspartic acid; C098885:dystrobrevin; D005047:Etoposide