Studies on the Mechanism of maturation and axonal transport of herpes simplex virus.

1.我们在单纯疱疹病毒中产生了UL 51缺失突变体（FDL 51），以揭示UL 51蛋白的功能。与野生型病毒相比，突变体噬斑的尺寸要小得多，FDL 51的最大滴度降低了近100倍。在FDL 51感染的细胞中，在核周间隙中观察到大量的包膜核衣壳，但很少检测到包膜成熟的病毒粒子，这表明UL 51蛋白参与了HSV-1粒子在初始纯化步骤下游的成熟和排出. UL 56是一种尾锚定的II型膜蛋白，与KIF 1A相关，KIF 1A是一种神经元特异性驱动蛋白，参与突触囊泡前体的轴突运输。UL 56蛋白还与UL 11蛋白相互作用，后者可能参与了核衣壳的形成和排出.在不存在US 3蛋白激酶（PK）的情况下，UL 46片段蛋白非常不稳定，更容易降解。UL 46蛋白在US 3缺陷型病毒的细胞外病毒粒子中检测不到。我们还发现US 3 PK可以直接磷酸化UL 46蛋白以及UL 34、UL 31、ICP 22和US9.4。我们已经确定了HF 10基因组的完整核苷酸序列，HF 10是一种高度减毒的非神经侵袭性HSV-1，并发现HF 10缺乏UL 56，UL 55，UL 43，UL49.5和潜伏相关转录本（LAT）的表达。

1. We generated UL51-null mutants(FDL51) in HSV to uncover the function of UL51 protein The mutant plaques were much smaller in size and maximal titers of FDL51 were reduced nearly 100-fold compared to wild-type virus. In FDL51-infected cells, a large number of envelope nucleocapsids were observed in the perinuclear space, but envelope mature virions were rarely detectable, suggesting that UL51 protein is involved in the maturation and egress of HSV-1 particles downstream of the initial envelopment step.2. UL56, a tail-anclcored type II membrane protein, associated with KIF1A, a neuron-specific kinesin involved in the axonal transport of synaptic vesicle precursors. UL56 protein also interacted with UL11 protein which is though to be involved in nuclear capsid envelopment and egress.3. In the absence of US3 protein kinase(PK), UL46 tegment protein was quite unstable, being much more susceptible to degradation. UL46 protein was undetectable in the extracellular virions of US3-deficient virus. We also found that US3PK can directy phosphorylate UL46 protein as well as UL34, UL31, ICP22, and US9.4. We have determined the complete nucleotide sequences of the genome of HF10, a highly attenuated, non-neuroinvasive HSV-1, and found that HF10 lacks the expression of UL56, UL55, UL43, UL49.5 and latency-associated transcripts(LAT).