Department of Developmental Biology, Institute of Biochemistry, LT-08662 Vilnius, Lithuania.
Mol Biol Rep. 2011 Jun;38(5):3001-11. doi: 10.1007/s11033-010-9965-9. Epub 2010 Jan 29.
Dystrobrevins (DBs) bind directly to dystrophin and are prominent components of the dystrophin-associated protein complex (DAPC) that links the cytoskeleton to the extracellular matrix. They are involved in brain development, synapse formation and plasticity, as well as water and ion homeostasis. However, the role of DB in non-muscular cells is not clear. In this study, we show that different α-dystrobrevin isoforms are present in promyelocytic leukemia (NB4) cells. Only the biggest α-dystrobrevin isoform (DB-α), which can be important for its function, was expressed in the membrane fraction of NB4 cells; the other α-DB isoforms were found in the hydrophilic cell fractions. Employing the immunoprecipitation and mass spectrometry, we identified novel α-DB-interacting proteins involved in cytoskeleton reorganization (actin, tropomyosin, gelsolin, tubulin) and signal transduction process (stathmin, prohibitin, RIBA) during proliferation and differentiation of NB4 cells. Our results suggest that α-DB isoforms play a central role in cytoskeleton reorganization via their multiple interactions with actin and actin-associating proteins and may participate in signal transduction process during NB4 cell granulocytic differentiation via directly and non directly associated proteins.
肌萎缩蛋白相关糖蛋白复合物(DAPC)将细胞骨架与细胞外基质连接在一起,而 dystrobrevins(DBs)直接与肌萎缩蛋白结合,是其重要组成部分。它们参与脑发育、突触形成和可塑性以及水和离子稳态。然而,DB 在非肌肉细胞中的作用尚不清楚。在这项研究中,我们表明不同的α-肌萎缩蛋白相关糖蛋白复合物(DAPC)在 promyelocytic leukemia(NB4)细胞中存在。只有最大的α-肌萎缩蛋白相关糖蛋白复合物(DAPC)(DB-α)在 NB4 细胞的膜部分表达;其他α-DB 异构体存在于亲水性细胞部分。通过免疫沉淀和质谱分析,我们鉴定了在 NB4 细胞增殖和分化过程中参与细胞骨架重排(肌动蛋白、原肌球蛋白、凝胶蛋白、微管蛋白)和信号转导过程(stathmin、prohibitins、RIBA)的新型α-DB 相互作用蛋白。我们的结果表明,α-DB 异构体通过与肌动蛋白和肌动蛋白相关蛋白的多种相互作用在细胞骨架重排中发挥核心作用,并可能通过直接和非直接相关蛋白参与 NB4 细胞粒细胞分化过程中的信号转导过程。