Cell Biology Laboratories, School of Biochemistry, Faculty of Life Sciences, University of Bristol, Bristol, UK.
School of Physiology, Pharmacology and Neuroscience, Faculty of Life Sciences, University of Bristol, Bristol, UK.
J Cell Biol. 2021 Jun 7;220(6). doi: 10.1083/jcb.202005166. Epub 2021 May 4.
Knockout of the golgin giantin leads to skeletal and craniofacial defects driven by poorly studied changes in glycosylation and extracellular matrix deposition. Here, we sought to determine how giantin impacts the production of healthy bone tissue by focusing on the main protein component of the osteoid, type I collagen. Giantin mutant zebrafish accumulate multiple spontaneous fractures in their caudal fin, suggesting their bones may be more brittle. Inducing new experimental fractures revealed defects in the mineralization of newly deposited collagen as well as diminished procollagen reporter expression in mutant fish. Analysis of a human giantin knockout cell line expressing a GFP-tagged procollagen showed that procollagen trafficking is independent of giantin. However, our data show that intracellular N-propeptide processing of pro-α1(I) is defective in the absence of giantin. These data demonstrate a conserved role for giantin in collagen biosynthesis and extracellular matrix assembly. Our work also provides evidence of a giantin-dependent pathway for intracellular procollagen processing.
高尔基巨蛋白的敲除导致糖基化和细胞外基质沉积变化引起的骨骼和颅面缺陷。在这里,我们通过关注成骨细胞外基质的主要蛋白质成分 I 型胶原,来研究高尔基巨蛋白如何影响健康骨组织的产生。高尔基巨蛋白突变的斑马鱼在其尾鳍上积累了多个自发性骨折,这表明它们的骨骼可能更脆弱。诱导新的实验性骨折显示,新沉积的胶原的矿化存在缺陷,并且突变鱼中的前胶原报告基因表达减少。分析表达 GFP 标记前胶原的人类高尔基巨蛋白敲除细胞系表明,前胶原的运输不依赖于高尔基巨蛋白。然而,我们的数据表明,在缺乏高尔基巨蛋白的情况下,前α1(I)的 N-前肽的细胞内加工是有缺陷的。这些数据表明高尔基巨蛋白在胶原生物合成和细胞外基质组装中具有保守作用。我们的工作还为细胞内前胶原加工的高尔基巨蛋白依赖性途径提供了证据。