Mechanism and regulation of protein kinase functions in HSV nuclear egress

HSV核出口中蛋白激酶功能的机制和调控

基本信息

批准号：
10088400
负责人：
RICHARD J ROLLER
金额：
$ 19.31万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-24 至 2022-12-31
项目状态：
已结题

项目摘要

Alphaherpesviruses encode two proteins kinases that are critical regulators of pathogenesis, pUS3 and pUL13. pUS3 has multiple functions in infection including facilitation of nuclear egress, protection from apoptosis, promotion of viral protein synthesis, and inhibition of antigen presentation to the immune system. The regulation of the activity of pUS3 is, therefore, of great interest, both as a tool for understanding the molecular biology of herpesvirus infections, and as an avenue for exploring antiviral therapies based on inhibition of its various activities. pUS3 kinase activity has been reported to be regulated both by autophosphorylation and by phosphorylation by another herpesvirus-encoded kinase, pUL13. Intriguingly, regulation of pUS3 activity by pUL13 appears to affect some pUS3 functions, but not others. Specifically, it appears to be necessary for at least some of the activities of pUS3 in egress of virus capsids from the nucleus, but not for protection of infected cells from some pro-apoptotic stimuli (). These observations highlight two very significant gaps in our understanding of the interaction between pUL13 and pUS3. First, it is unclear which pUS3 functions are regulated by pUL13, and what distinguished UL13-dependent from UL13-independent functions. We propose the simple hypothesis that phosphorylation of pUS3 by pUL13 specifically regulates its nuclear functions by regulating its access to the nucleus. Second, the mechanism and functional significance of pUL13 regulation of pUS3 is unclear. It is known that pUL13 phosphorylates pUS3, but it is not clear whether this phosphorylation is necessary for pUL13 regulation of pUS3. A recently published proteomic analysis of HSV infected cells suggests that pUL13 phosphorylates pUS3 at S139. We will use a genetic approach to test the hypothesis that S139 is the critical residue for regulation of pUS3 by pUL13, and then to test the significance of S139 phosphorylation for viral growth and spread.

α-掌病毒编码两种蛋白质激酶，这些激酶是发病机理的关键调节剂PUS3和PUL13。 PUS3在感染中具有多个功能，包括促进核出口，免受凋亡，促进病毒蛋白合成以及抑制对免疫系统的抗原表现。因此，PUS3活性的调节引起了人们的极大关注，既是理解疱疹病毒感染的分子生物学的工具，又是基于抑制其各种活动的抗病毒药疗法的途径。据报道，PUS3激酶活性是通过自磷酸化和通过另一种疱疹病毒编码激酶PUL13的磷酸化来调节的。有趣的是，PUL13对PUS3活性的调节似乎会影响某些PUS3功能，但没有影响其他PUS3。具体而言，对于至少某些PUS3在细胞核中流出的PUS3活性似乎是必要的，但对于保护感染细胞免受某些促凋亡刺激（）（）（）（）。这些观察结果突出了我们对PUL13和PUS3之间相互作用的理解时的两个非常明显的差距。首先，尚不清楚哪些PUS3函数由pUL13调节，以及与UL13独立函数区别的ul13依赖性。我们提出了这样一个简单的假设，即PUL13对PUS3的磷酸化特异性调节其核功能通过调节其进入核的访问。其次，PUS3调控的机理和功能意义尚不清楚。众所周知，PUL13磷酸化PUS3，但尚不清楚这种磷酸化是否对于PUS3的PUL13调节是必需的。最近发表的HSV感染细胞的蛋白质组学分析表明，PUL13在S139处磷酸化PUS3。我们将使用一种遗传方法来检验以下假设：S139是通过PUL13调节PUS3的关键残基，然后测试S139磷酸化对病毒生长和扩散的显着性。