MMP-9 facilitates selective proteolysis of the histone H3 tail at genes necessary for proficient osteoclastogenesis.
MMP-9 facilitates selective proteolysis of the histone H3 tail at genes necessary for proficient osteoclastogenesis.
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DOI：
10.1101/gad.268714.115
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发表时间：
2016-01-15
影响因子：
10.5
通讯作者：
An W
中科院分区：
文献类型：
--
作者：
Kim K;Punj V;Kim JM;Lee S;Ulmer TS;Lu W;Rice JC;An W
金等人。发现组蛋白 H3 N 末端尾部 (H3NT) 蛋白水解选择性地靶向一小群基因的转录起始位点附近，并且大多数 H3NT 切割的基因在破骨细胞生成过程中表现出显着的表达变化。破骨细胞生成的主要 H3NT 蛋白酶是基质金属蛋白酶 9 (MMP-9)。尽管在哺乳动物分化过程中经常观察到组蛋白 H3 N 末端尾部 (H3NT) 的有限蛋白水解，但针对 H3NT 蛋白水解的特定基因组位点以及 H3NT 裂解的功能意义仍然很大程度上未知。在这里，我们报告了第一种识别和检查哺乳动物中 H3NT 切割区域的方法，称为乙酰化染色质染色质免疫沉淀 (ChIP) (ChIPac)。通过将 ChIPac 结合深度测序（ChIPac-seq）应用于已建立的破骨细胞分化细胞模型，我们发现 H3NT 蛋白水解选择性地靶向一小群基因的转录起始位点附近，并且大多数 H3NT 裂解的基因在破骨细胞生成过程中表现出显着的表达变化。我们还发现破骨细胞生成的主要 H3NT 蛋白酶是基质金属蛋白酶 9 (MMP-9)。与其他已知的 H3NT 蛋白酶相比，MMP-9 主要在体外和细胞内裂解 H3K18-Q19。此外，我们的结果支持 CBP/p300 介导的 H3K18 乙酰化作为体外和破骨细胞生成过程中 H3NT 裂解位点 MMP-9 H3NT 蛋白酶活性的中心调节剂。重要的是，我们发现H3NT蛋白水解作用的废除会损害破骨细胞生成基因的激活，同时导致破骨细胞分化缺陷。我们的集体结果支持 MMP-9 依赖性 H3NT 蛋白水解在调节熟练破骨细胞生成所需的基因途径中的必要性。
Kim et al. discovered that histone H3 N-terminal tail (H3NT) proteolysis is selectively targeted near transcription start sites of a small group of genes and that most H3NT-cleaved genes displayed significant expression changes during osteoclastogenesis. The principal H3NT protease of osteoclastogenesis is matrix metalloproteinase 9 (MMP-9). Although limited proteolysis of the histone H3 N-terminal tail (H3NT) is frequently observed during mammalian differentiation, the specific genomic sites targeted for H3NT proteolysis and the functional significance of H3NT cleavage remain largely unknown. Here we report the first method to identify and examine H3NT-cleaved regions in mammals, called chromatin immunoprecipitation (ChIP) of acetylated chromatin (ChIPac). By applying ChIPac combined with deep sequencing (ChIPac-seq) to an established cell model of osteoclast differentiation, we discovered that H3NT proteolysis is selectively targeted near transcription start sites of a small group of genes and that most H3NT-cleaved genes displayed significant expression changes during osteoclastogenesis. We also discovered that the principal H3NT protease of osteoclastogenesis is matrix metalloproteinase 9 (MMP-9). In contrast to other known H3NT proteases, MMP-9 primarily cleaved H3K18-Q19 in vitro and in cells. Furthermore, our results support CBP/p300-mediated acetylation of H3K18 as a central regulator of MMP-9 H3NT protease activity both in vitro and at H3NT cleavage sites during osteoclastogenesis. Importantly, we found that abrogation of H3NT proteolysis impaired osteoclastogenic gene activation concomitant with defective osteoclast differentiation. Our collective results support the necessity of MMP-9-dependent H3NT proteolysis in regulating gene pathways required for proficient osteoclastogenesis.