Laboratory of Cellular and Molecular Neurobiology, Hellenic Pasteur Institute, Athens, Greece.
PLoS One. 2013 Nov 28;8(11):e82172. doi: 10.1371/journal.pone.0082172. eCollection 2013.
BM88/Cend1 is a neuronal-lineage specific modulator with a pivotal role in coordination of cell cycle exit and differentiation of neuronal precursors. In the current study we identified the signal transduction scaffolding protein Ran-binding protein M (RanBPM) as a BM88/Cend1 binding partner and showed that BM88/Cend1, RanBPM and the dual specificity tyrosine-phosphorylation regulated kinase 1B (Dyrk1B) are expressed in mouse brain as well as in cultured embryonic cortical neurons while RanBPM can form complexes with either of the two other proteins. To elucidate a potential mechanism involving BM88/Cend1, RanBPM and Dyrk1B in cell cycle progression/exit, we transiently co-expressed these proteins in mouse neuroblastoma Neuro 2a cells. We found that the BM88/Cend1-dependent or Dyrk1B-dependent down-regulation of cyclin D1 is reversed following their functional interaction with RanBPM. More specifically, functional interaction of RanBPM with either BM88/Cend1 or Dyrk1B stabilizes cyclin D1 in the nucleus and promotes 5-bromo-2'-deoxyuridine (BrdU) incorporation as a measure of enhanced cell proliferation. However, the RanBPM-dependent Dyrk1B cytosolic retention and degradation is reverted in the presence of Cend1 resulting in cyclin D1 destabilization. Co-expression of RanBPM with either BM88/Cend1 or Dyrk1B also had a negative effect on Neuro 2a cell differentiation. Our results suggest that functional interactions between BM88/Cend1, RanBPM and Dyrk1B affect the balance between cellular proliferation and differentiation in Neuro 2a cells and indicate that a potentially similar mechanism may influence cell cycle progression/exit and differentiation of neuronal precursors.
BM88/Cend1 是一种神经元谱系特异性调节剂,在协调细胞周期退出和神经元前体细胞分化方面起着关键作用。在本研究中,我们鉴定了信号转导支架蛋白 Ran 结合蛋白 M(RanBPM)作为 BM88/Cend1 的结合伴侣,并表明 BM88/Cend1、RanBPM 和双特异性酪氨酸磷酸化调节激酶 1B(Dyrk1B)在小鼠大脑中以及在培养的胚胎皮质神经元中表达,而 RanBPM 可以与这两种蛋白质中的任何一种形成复合物。为了阐明 BM88/Cend1、RanBPM 和 Dyrk1B 在细胞周期进程/退出中的潜在机制,我们在小鼠神经母细胞瘤 Neuro 2a 细胞中瞬时共表达这些蛋白质。我们发现,BM88/Cend1 依赖性或 Dyrk1B 依赖性下调 cyclin D1 可在其与 RanBPM 的功能相互作用后逆转。更具体地说,RanBPM 与 BM88/Cend1 或 Dyrk1B 的功能相互作用稳定核内 cyclin D1,并促进 5-溴-2'-脱氧尿苷(BrdU)掺入作为增强细胞增殖的衡量标准。然而,在 Cend1 存在的情况下,RanBPM 依赖性 Dyrk1B 细胞质保留和降解被逆转,导致 cyclin D1 不稳定。RanBPM 与 BM88/Cend1 或 Dyrk1B 的共表达也对 Neuro 2a 细胞分化产生负面影响。我们的结果表明,BM88/Cend1、RanBPM 和 Dyrk1B 之间的功能相互作用影响 Neuro 2a 细胞中细胞增殖和分化之间的平衡,并表明类似的潜在机制可能影响神经元前体细胞的细胞周期进程/退出和分化。