Vázquez-Abad D, Carson J H, Rothfield N
Division of Rheumatic Diseases, Department of Medicine, University of Connecticut School of Medicine, Farmington, CT 06030, USA.
Cell Tissue Res. 1996 Dec;286(3):487-91. doi: 10.1007/s004410050718.
The present study was designed to determine the subcellular localization of histidyl-tRNA synthetase (Jo-1) in human laryngeal epithelial carcinoma cell line (HEp-2 cells). Indirect immunofluorescence using commercial HEp-2 cells with human serum and human-affinity-purified anti-Jo-1 antibodies was performed using confocal microscopy. Anti-histidyl-tRNA-synthetase-positive sera showed distinct nuclear and cytoplasmic granular staining in HEp-2 cells. Affinity purified anti-Jo-1 produced an identical pattern to the whole serum, whereas the serum fraction that did not bind to the affinity column was negative by immunofluorescence on HEp-2 cells. Two commercial human anti-Jo-1-positive control sera and seven anti-Jo-1-positive sera from patients with myositis reproduced the nuclear and cytoplasmic granular pattern. We conclude that Jo-1 is present in cytoplasm and in intact nuclei from HEp-2 cells. The presence of tRNA synthetases in intact nuclei suggests that they have an unsuspected function in the nucleus.
本研究旨在确定组氨酰 - tRNA合成酶（Jo - 1）在人喉上皮癌细胞系（HEp - 2细胞）中的亚细胞定位。使用共聚焦显微镜，对商业化的HEp - 2细胞、人血清和人亲和纯化的抗Jo - 1抗体进行间接免疫荧光检测。抗组氨酰 - tRNA合成酶阳性血清在HEp - 2细胞中显示出明显的细胞核和细胞质颗粒状染色。亲和纯化的抗Jo - 1产生的染色模式与全血清相同，而未与亲和柱结合的血清组分在HEp - 2细胞上的免疫荧光检测为阴性。两种商业化的人抗Jo - 1阳性对照血清和来自肌炎患者的七种抗Jo - 1阳性血清重现了细胞核和细胞质颗粒状模式。我们得出结论，Jo - 1存在于HEp - 2细胞的细胞质和完整细胞核中。完整细胞核中存在tRNA合成酶表明它们在细胞核中具有未被怀疑的功能。
