Early interactions of DNA viruses and host

项目描述 人巨细胞病毒（HCMV）感染对免疫功能低下的个体具有严重后果， 例如艾滋病和器官移植患者以及先天感染的新生儿。有症状的新生儿， 12%的人死亡，许多幸存者患有精神发育迟滞，视力丧失和感觉神经性耳聋。电流 针对病毒DNA复制早期事件的HCMV干扰策略既不被允许， 用于治疗受感染的胎儿是可行的，并且可能导致耐药性。我们建议调查 CMV感染周期的立即早期阶段，以确定新的潜在治疗靶点。即刻早期 蛋白1和2（IE1，IE2）对于HCMV的复制成功和再活化是必需的， 延迟。我们假设：1.受调控的选择性剪接决定了多步骤中的第二步 从潜伏期重新激活的过程，以及2. IE1抵消内在的核防御，如基因沉默 和诱导细胞凋亡，因此阻断IE蛋白功能将大大减少或完全消除病毒性肝炎。 复制的为了验证这些假设，我们将研究人类和小鼠CMV的主要IE蛋白 以建立功能同源性。具体来说，我们将确定：1。拼接如何决定 数量和时间的主要立即早期转录单位，2。小鼠模型是否可以诱导 表达立即早期蛋白可以阐明多步再激活序列中的早期步骤 以及IE1和IE2的细胞特异性表达，以及3. IE1如何对抗基于基因沉默的抗病毒药物 宿主细胞的防御，目的是决定它是否是阻断裂解的合理靶标， 再活化级联来自这些分析的信息将改变CMV干扰选项的范例 远离复制性事件而朝向感染的早期事件。

Project Description Human cytomegalovirus (HCMV) infection has serious consequences for immunocompromised individuals, such as AIDS and organ transplant patients and congenitally infected newborns. Of symptomatic newborns, 12% die and many of the survivors suffer mental retardation, vision loss, and sensorineural deafness. Current HCMV interference strategies targeting early events in viral DNA replication are neither permissible nor feasible for treating the infected fetus, and may lead to drug resistance. We propose to investigate the immediate-early stage of the CMV infectious cycle to identify new potential targets of therapy. The immediateearly proteins 1 and 2 (IE1, IE2) are essential for the replicative success and reactivation of HCMV from latency. We hypothesize that: 1. Regulated alternative splicing determines the second step in the multi-step process of reactivation from latency, and 2. IE1 counteracts intrinsic nuclear defenses such as gene silencing and induction of apoptosis, so that a block in IE protein function will greatly reduce or totally abrogate viral replication. To test these hypotheses, we will investigate the major IE proteins of both human and mouse CMV in parallel to establish functional homology. Specifically, we will determine: 1. How splicing determines the amount and timing of the major immediate early transcription unit, 2. Whether a mouse model inducibly expressing the immediate early proteins can elucidate the early steps in the multi-step reactivation sequence and cell-specific expression of IE1 and IE2, and 3. How IE1 counters the gene silencing-based antiviral defenses of the host cell with the goal of deciding whether it is a reasonable target to block the lytic and reactivation cascade. Information from these analyses will shift the paradigm for CMV interference options away from replicative events and toward earlier events in the infection.