Acetylation of NDPK-D Regulates Its Subcellular Localization and Cell Survival.
Acetylation of NDPK-D Regulates Its Subcellular Localization and Cell Survival.
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DOI：
10.1371/journal.pone.0139616
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yamashita T
中科院分区：
文献类型：
--
作者：
Fujita Y;Fujiwara K;Zenitani S;Yamashita T
核苷二磷酸激酶（NDPK）是一种普遍存在的酶，它催化γ-磷酸在二磷酸和三磷酸核苷之间的可逆磷酸转移。NDPK- d （Nm23-H4）是NDPK家族中唯一具有线粒体靶向序列的成员。尽管NDPK-D在发育中的中枢神经系统中高表达，但其功能尚不清楚。在这项研究中，我们发现NDPK-D敲低可诱导神经母细胞瘤细胞和小鼠皮层的凋亡，这表明NDPK-D是神经元存活所必需的。通过酵母双杂交筛选，我们发现NDPK-D是NAD+依赖性组蛋白去乙酰化酶SIRT1的结合伙伴。NDPK-D与SIRT1共定位，通过共免疫沉淀证实了这些分子的关联。抑制SIRT1增加NDPK-D的乙酰化。NDPK-D与SIRT1的过表达，或NDPK-D中乙酰化赖氨酸残基的突变，增加了其核积累。此外，NDPK-D乙酰化模拟突变体增加了N1E-115细胞的凋亡。我们的数据表明，乙酰化调节NDPK-D在细胞核和细胞质之间的穿梭，NDPK-D乙酰化增加导致细胞凋亡。
Nucleoside diphosphate kinases (NDPK) are ubiquitous enzymes that catalyze the reversible phosphotransfer of γ-phosphates between di- and triphosphonucleosides. NDPK-D (Nm23-H4) is the only member of the NDPK family with a mitochondrial targeting sequence. Despite the high expression of NDPK-D in the developing central nervous system, its function remains to be determined. In this study, we show that NDPK-D knockdown induces apoptosis in neuroblastoma cells as well as in mouse cortex, suggesting that NDPK-D is required for neuronal survival. We identified NDPK-D as a binding partner of NAD+-dependent histone deacetylase, SIRT1, by yeast two-hybrid screening. NDPK-D co-localized with SIRT1, and the association of these molecules was confirmed by co-immunoprecipitation. Inhibition of SIRT1 increases the acetylation of NDPK-D. Overexpression of NDPK-D along with SIRT1, or mutation in the acetylated lysine residues in NDPK-D, increases its nuclear accumulation. Furthermore, the NDPK-D acetylation-mimic mutant increased apoptosis in N1E-115 cells. Our data demonstrate that acetylation regulates the shuttling of NDPK-D between nucleus and cytoplasm, and increased acetylation of NDPK-D causes apoptosis.