Regulation of the G2/M phase of the cell cycle by sperm associated antigen 8 (SPAG8) protein
Regulation of the G2/M phase of the cell cycle by sperm associated antigen 8 (SPAG8) protein
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DOI：
10.1002/cbf.1574
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发表时间：
2009-07-01
影响因子：
3.6
通讯作者：
Wang, Lin-Fang
中科院分区：
文献类型：
--
作者：
Li, Rong;Tang, Xiao-Ling;Wang, Lin-Fang
精子相关抗原8(SPAG8)是一种在男性生殖细胞分化过程中产生的睾丸特异蛋白，它是利用从一名不育妇女血清中发现的抗体从人类睾丸表达文库中分离出来的。发现它与微管有密切的功能关系。在本研究中，我们建立了稳定表达SPAG8的CHO-K1细胞系。免疫荧光共聚焦显微镜显示SPAG8在早期集中在微管组织中心(MTOC)。随着细胞进入中期，它与纺锤体上的α-微管蛋白共定位。在发育后期，星形微管和中间带均可检测到。胞质分裂后，SPAG8重新定位于MTOC。同时，流式细胞仪分析发现，SPAG8延长了稳定表达SPAG8的CHO-K1细胞的G2/M期。四甲基偶氮唑盐(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)比色法显示SPAG8对稳定细胞的增殖有抑制作用。SPAG8可能通过改变CDC2上Tyr15的磷酸化水平而参与细胞周期的调控。这些结果提示，SPAG8可能在精子发生过程中参与细胞分裂。版权所有(C)2009 John Wiley&Sons，Ltd.
Sperm associated antigen 8 (SPAG8), a testis-specific protein produced during male germ cell differentiation, was isolated from a human testis expression library using antibodies found in the serum obtained from an infertile woman. It was found to have a close functional relationship with microtubules. In this study, we generated a stably expressing SPAG8 CHO-K1 cell line. Immunofluorescence confocal microscopy showed that SPAG8 was concentrated at the microtubule-organizing center (MTOC) during prophase. As the cells progressed into metaphase, it co-localized with a-tubulin on the spindle. In anaphase, it was detected on both astral microtubules and mid-zone. Following cytokinesis, SPAG8 resumed its localization on the MTOC. Meanwhile, flow cytometry analysis found that SPAG8 prolonged the G2/M phase of CHO-K1 cells stably expressing SPAG8. Furthermore, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that SPAG8 inhibited the proliferation of the stable cells. SPAG8 might be involved in the regulation of cell cycle by changing the phosphorylation level of Tyr15 on cdc2. These results suggest that SPAG8 might play a role in cell division during spermatogenesis. Copyright (C) 2009 John Wiley & Sons, Ltd.