BMP-1-induced GBA1 nuclear accumulation provokes CCN2 mRNA expression via importin-β-mediated nucleocytoplasmic pathway
BMP-1-induced GBA1 nuclear accumulation provokes CCN2 mRNA expression via importin-β-mediated nucleocytoplasmic pathway
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DOI：
10.1007/s12079-023-00740-3
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发表时间：
2023-03-27
影响因子：
4.1
通讯作者：
Tani-Ishii, Nobuyuki
中科院分区：
文献类型：
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作者：
Muromachi, Koichiro;Nakano, Rei;Tani-Ishii, Nobuyuki
骨形态发生蛋白(BMP)-1由牙本质-牙髓复合体中的成牙本质细胞表达。尽管BMP-1在启动矿化的各种蛋白质和酶的预形体成熟过程中的功能效应已经被广泛观察到，但BMP-1如何影响细胞分子仍不清楚。我们对BMP-1改变的糖链进行了全面的分析，并通过糖代谢的方法鉴定了人牙髓细胞(HDPC)中的糖蛋白。在BMP-1存在的情况下，凝集素微阵列分析和凝集素探针印迹显示，hDPC的不溶性部分的α2，6-唾液酸化显著减弱。用凝集素柱纯化的α-2，6-唾液酸糖蛋白进行了质谱分析，鉴定了6种蛋白质。其中，在BMP-1存在的情况下，葡萄糖神经酰胺酶(GBA1)聚集在hDPC的细胞核中。此外，BMP-1诱导的细胞通讯网络因子(CCN)2的表达在转GBA1 siRNA的细胞中受到显著抑制，CCN是成骨/软骨形成的标志。此外，Importin-beta介导的核进口的有效抑制剂Importazole显著抑制BMP-1诱导的GBA1核聚集和BMP-1诱导的CCN2mRNA表达。因此，BMP-1通过还原α2，6-唾液酸促进GBA1在细胞核中的积累，这可能通过Importin-β介导的hDPC核输入途径参与CCN2基因的转录调控。我们的结果为BMP-1-GBA1-CCN2轴在牙科/颅面疾病的发育、组织重塑和病理中的作用提供了新的见解。
Bone morphogenetic protein (BMP)-1 is expressed by odontoblasts in the dentin-pulp complex. Although the functional effects of BMP-1 on the maturation of various preforms of proteins and enzymes involved in initiating mineralization have been widely observed, how BMP-1 affects cellular molecules remains unknown. We performed a comprehensive analysis of BMP-1-altered glycome profiles and subsequent assays to identify the target glycoproteins in human dental pulp cells (hDPCs) by a glycomic approach. In the presence of BMP-1, a lectin microarray analysis and lectin-probed blotting showed that alpha 2,6-sialylation was significantly attenuated in insoluble fractions from hDPCs. Six proteins were identified by a mass spectrometry analysis of alpha 2,6-sialylated glycoproteins purified using a lectin column. Among them, glucosylceramidase (GBA1) was found to accumulate in the nuclei of hDPCs in the presence of BMP-1. Moreover, BMP-1-induced cellular communication network factor (CCN) 2 expression, which is well known as the osteogenesis/chondrogenesis marker, was significantly suppressed in the cells transfected with GBA1 siRNA. Furthermore, importazole, a potent inhibitor of importin-beta-mediated nuclear import significantly suppressed BMP-1-induced GBA1 nuclear accumulation and BMP-1-induced CCN2 mRNA expression, respectively. Thus, BMP-1 facilitates the accumulation of GBA1 in the nucleus through the reduction of alpha 2,6-sialic acid, which potentially contributes to the transcriptional regulation of the CCN2 gene via importin-beta-mediated nuclear import pathway in hDPCs. Our results offer new insights into the role of the BMP-1-GBA1-CCN2 axis in the development, tissue remodeling, and pathology of dental/craniofacial diseases.[GRAPHICS].