...
首页> 外文期刊>Cell cycle >Characterization and functional aspects of human ninein isoforms that regulated by centrosomal targeting signals and evidence for docking sites to direct gamma-tubulin.
【24h】

Characterization and functional aspects of human ninein isoforms that regulated by centrosomal targeting signals and evidence for docking sites to direct gamma-tubulin.

机译:受中心体靶向信号调控的人类九蛋白同工型的表征和功能方面,以及直接对接γ-微管蛋白的停靠位点的证据。

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       

摘要

The functions of centrosomal protein ninein may be involved in microtubule minus end capping, centriole positioning, protein anchoring and microtubule nucleation, but the true physiological function of various human hNinein isoforms remains to be determined. Here we describe the identification of four diverse CCII-termini of human hNinein isoforms, including a novel isoform 6, by differential expression in a tissue-specific manner. These hNinein isoforms exhibit centrosomal (concentrated) and noncentrosomal (aggregated) localization when GFP-tagged fusion proteins are expressed transiently in mammalian cells. In a kinase assay, we show that the CCII region of hNinein provides a differential phosphorylation site by GSK3beta. In addition, our data indicate that either N-terminal or CCIIZ domain disruption may cause hNinein conformational change which recruits gamma-tubulin to centrosomal or noncentrosomal hNinein-containing sites, implying that the gamma-tubulin localization may be hNinein-dependent. Further, our RNA interference experiment against all hNinein isoforms caused a significant decrease in the gamma-tubulin signal in the centrosome. In domain swapping, we clearly show that the CCIIX-CCIIY region provides docking sites for gamma-tubulin. Moreover, our data also show that nucleation of microtubules from the centrosome is significantly affected by the presence of either the full -length hNinein or CCIIX-CCIIY region overexpression. Taken together, these results show that the centrosomal targeting signals of hNinein have a role not only in regulating hNinein conformation, resulting in localization change, but also provide docking sites to recruit gamma-tubulin at centrosomal and noncentrosomal sites.
机译:中心体蛋白ninein的功能可能涉及微管减去末端封端,中心蛋白定位,蛋白锚定和微管成核,但是各种人hNinein亚型的真正生理功能仍有待确定。在这里我们描述了人类hNinein亚型的四个不同CCII末端的鉴定,包括通过组织特异性方式的差异表达,包括一个新的亚型6。当在哺乳动物细胞中瞬时表达GFP标记的融合蛋白时,这些hNinein亚型表现出中心体(集中)和非中心体(聚集)定位。在激酶测定中,我们显示hNinein的CCII区通过GSK3beta提供了一个不同的磷酸化位点。此外,我们的数据表明,N末端或CCIIZ结构域破坏都可能引起hNinein构象变化,从而将γ-微管蛋白募集到含hNinein的中心体或非中心体位点,这意味着γ-微管蛋白的定位可能是hNinein依赖性的。此外,我们针对所有hNinein亚型的RNA干扰实验导致中心体中的γ-微管蛋白信号显着降低。在域交换中,我们清楚地表明CCIIX-CCIIY地区为γ-微管蛋白提供了对接位点。此外,我们的数据还表明,全长hNinein或CCIIX-CCIIY区过表达的存在显着影响了来自中心体的微管成核。综上,这些结果表明,hNinein的中心体靶向信号不仅在调节hNinein构象,导致定位改变方面起作用,而且还提供了在中心体和非中心体位点募集γ-微管蛋白的停靠位点。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号