Nuclear localization of DMP1 proteins suggests a role in intracellular signaling.
Nuclear localization of DMP1 proteins suggests a role in intracellular signaling.
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DOI：
10.1016/j.bbrc.2012.07.037
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发表时间：
2012-08-03
影响因子：
3.1
通讯作者：
Lu Y
中科院分区：
文献类型：
--
作者：
Siyam A;Wang S;Qin C;Mues G;Stevens R;D'Souza RN;Lu Y
牙本质基质蛋白 1 (DMP1) 在成牙本质细胞和成骨细胞/骨细胞中高度表达，在牙齿和骨矿化以及磷酸盐稳态中发挥重要作用。 DMP1除了在细胞外基质中发挥作用外，是否还能进入细胞核并发挥转录因子的作用，目前尚有争议。为了更好地了解其功能，我们检查了 C3H10T1/2 间充质细胞、MC3T3-E1 前成骨细胞和 17IIA11 成牙本质细胞样细胞中内源性和外源性 DMP1 的核定位。 RT-PCR 分析显示内源性 Dmp1 在所有三种细胞系中表达，而蛋白质印迹分析检测到主要的 DMP1 蛋白条带，对应于分泌的全长 DMP1 的蛋白水解加工产生的 57 kDa C 端片段。免疫荧光染色表明，非同步细胞呈现出两个内源性 DMP1 定位于核或细胞质的亚群。此外，当用抗HA标签的抗体检查时，用表达HA标签的全长DMP1的构建体转染的细胞也显示出外源DMP1的核或细胞质定位。此外，核DMP1仅限于核质，但在核仁中不存在。总之，这些发现表明，除了作为牙本质和骨基质成分的作用外，DMP1 可能在细胞核中发挥调节作用。
Dentin matrix protein 1 (DMP1) is highly expressed in odontoblasts and osteoblasts/osteocytes and plays an essential role in tooth and bone mineralization and phosphate homeostasis. It is debatable whether DMP1, in addition to its function in the extracellular matrix, can enter the nucleus and function as a transcription factor. To better understand its function, we examined the nuclear localization of endogenous and exogenous DMP1 in C3H10T1/2 mesenchymal cells, MC3T3-E1 preosteoblast cells and 17IIA11 odontoblast-like cells. RT-PCR analyses showed the expression of endogenous Dmp1 in all three cell lines, while Western-blot analysis detected a major DMP1 protein band corresponding to the 57 kDa C-terminal fragment generated by proteolytic processing of the secreted full-length DMP1. Immunofluorescent staining demonstrated that non-synchronized cells presented two subpopulations with either nuclear or cytoplasmic localization of endogenous DMP1. In addition, cells transfected with a construct expressing HA-tagged full-length DMP1 also showed either nuclear or cytoplasmic localization of the exogenous DMP1 when examined with an antibody against the HA tag. Furthermore, nuclear DMP1 was restricted to the nucleoplasm but was absent in the nucleolus. In conclusion, these findings suggest that, apart from its role as a constituent of dentin and bone matrix, DMP1 might play a regulatory role in the nucleus.
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