Yamauchi Yohei, Kiriyama Kazuya, Kimura Hiroshi, Nishiyama Yukihiro
Department of Virology, Graduate School of Medicine, Nagoya University, Nagoya, Japan.
J Cell Sci. 2008 Jun 15;121(Pt 12):2087-96. doi: 10.1242/jcs.031450. Epub 2008 May 27.
The nuclear mitotic apparatus (NuMA) protein is a component of the nuclear matrix in interphase cells and an essential protein for the formation of mitotic spindle poles. We used herpes simplex virus (HSV), an enveloped DNA virus that replicates in the nucleus, to study the intra-nuclear dynamics of NuMA in infected cells. This study shows that NuMA is extensively modified following HSV infection, including phosphorylation of an unidentified site(s), and that it depends to an extent on viral DNA synthesis. Although NuMA is insoluble in uninfected interphase cells, HSV infection induced solubilisation and dynamic relocalisation of NuMA, whereupon the protein became excluded from viral replication compartments -- sites of virus transcription and replication. Live cell, confocal imaging showed that NuMA localisation dramatically changed from the early stages (diffusely nuclear, excluding nucleoli) to late stages of infection (central diminuition, but remaining near the inner nuclear peripheries). In addition, NuMA knockdown using siRNA suggested that NuMA is important for efficient viral growth. In summary, we suggest that NuMA is required for efficient HSV infection, and identify further areas of research that address how the virus challenges host cell barriers.
核有丝分裂装置(NuMA)蛋白是间期细胞中核基质的一个组成部分,也是有丝分裂纺锤体极形成所必需的蛋白。我们利用单纯疱疹病毒(HSV),一种在细胞核中复制的包膜DNA病毒,来研究感染细胞中NuMA的核内动力学。本研究表明,HSV感染后NuMA会发生广泛修饰,包括一个未确定位点的磷酸化,并且在一定程度上依赖于病毒DNA合成。虽然NuMA在未感染的间期细胞中不溶,但HSV感染会诱导NuMA溶解并动态重新定位,随后该蛋白被排除在病毒复制区室之外,病毒复制区室是病毒转录和复制的位点。活细胞共聚焦成像显示,NuMA的定位从感染早期(弥漫性核内,排除核仁)到感染后期(中央减少,但仍靠近核内周边)发生了显著变化。此外,使用小干扰RNA(siRNA)敲低NuMA表明,NuMA对病毒的有效生长很重要。总之,我们认为NuMA是HSV有效感染所必需的,并确定了进一步的研究领域,以解决病毒如何挑战宿主细胞屏障的问题。