Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, 701, Taiwan.
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, 701, Taiwan.
J Cell Biochem. 2018 Jan;119(1):314-326. doi: 10.1002/jcb.26180. Epub 2017 Jun 27.
Leucine-rich repeats and WD repeat domain containing protein 1 (LRWD1) is a testis-specific protein that mainly expressed in the sperm neck where centrosome is located. By using microarray analysis, LRWD1 is identified as a putative gene that involved in spermatogenesis. However, its role in human male germ cell development has not been extensively studied. When checking in the semen of patients with asthenozoospermia, teratozoospermia, and asthenoteratozoospermia, the level of LRWD1 in the sperm neck was significantly reduced with a defective neck or tail. When checking the sub-cellular localization of LRWD1 in the cells, we found that LRWD1 resided in the centrosome and its centrosomal residency was independent of microtubule transportation in NT2/D1, the human testicular embryonic carcinoma, cell line. Depletion of LRWD1 did not induce centrosome re-duplication but inhibited microtubule nucleation. In addition, the G1 arrest were observed in LRWD1 deficient NT2/D1 cells. Upon LRWD1 depletion, the levels of cyclin E, A, and phosphorylated CDK2, were reduced. Overexpression of LRWD1 promoted cell proliferation in NT2/D1, HeLa, and 239T cell lines. In addition, we also observed that autophagy was activated in LRWD1 deficient cells and inhibition of autophagy by chloroquine or bafilomycin A1 promoted cell death when LRWD1 was depleted. Thus, we found a novel function of LRWD1 in controlling microtubule nucleation and cell cycle progression in the human testicular embryonic carcinoma cells. J. Cell. Biochem. 119: 314-326, 2018. © 2017 Wiley Periodicals, Inc.
富含亮氨酸重复和 WD 重复域蛋白 1（LRWD1）是一种睾丸特异性蛋白，主要表达在中心体所在的精子颈部。通过使用微阵列分析，LRWD1 被鉴定为一个可能参与精子发生的基因。然而，其在人类男性生殖细胞发育中的作用尚未得到广泛研究。在检查弱精子症、畸形精子症和弱畸形精子症患者的精液时，LRWD1 在颈部的水平明显降低，伴有颈部或尾部缺陷。在检查 LRWD1 在细胞中的亚细胞定位时，我们发现 LRWD1 位于中心体，其中心体驻留独立于 NT2/D1（人睾丸胚胎癌）细胞系中的微管运输。LRWD1 的缺失不会诱导中心体重复复制，但会抑制微管的起始。此外，在 LRWD1 缺陷的 NT2/D1 细胞中观察到 G1 期阻滞。LRWD1 缺失后，细胞周期蛋白 E、A 和磷酸化 CDK2 的水平降低。LRWD1 的过表达促进了 NT2/D1、HeLa 和 239T 细胞系的细胞增殖。此外，我们还观察到自噬在 LRWD1 缺陷细胞中被激活，并且当 LRWD1 缺失时，用氯喹或巴弗洛霉素 A1 抑制自噬会促进细胞死亡。因此，我们发现了 LRWD1 在控制人睾丸胚胎癌细胞中微管起始和细胞周期进程中的新功能。J. Cell. Biochem. 119: 314-326, 2018. © 2017 Wiley Periodicals, Inc.
