MURINE GAMMAHERPESVIRUS 68 GENE EXPRESSION AND FUNCTION

描述(改编自调查人员的摘要)：主要目标 这项研究的特点是在体外表征一种细胞凋亡性疾病的调节 小鼠伽马疱疹病毒68株M11基因编码的病毒Bcl2同源物 (MHV-68)。M11开放阅读框位于病毒基因簇内 这可能与MHV-68的潜伏期相关。这个基因簇存在于 相关的横纹状病毒包括KSHV和疱疹病毒(HSV)，但 该簇中存在Bcl2同源基因是MHV-68所特有的。然而， KSHV、HVS和包括EBV在内的其他伽马疱疹病毒基因组也编码 与M11有潜在可比性的BCL-2同源蛋白。在 建议在体外研究重组病毒的构建和M11的表达 细胞系将被用于探索M11的功能。精心设计的MHV-68 利用野生型病毒进行重组体和常规表达研究 也有望提供对M11在 体外特定的细胞类型提供了可能在以下情况下追求的初步数据 在未来的活体研究中有必要超出当前提议的范围。 拟议研究的具体目标包括：1)分析M11的表达 和邻近基因在NIH/3T3细胞裂解感染过程中和在 小鼠B淋巴细胞和巨噬细胞感染的实验研究 2)M11在选定细胞中的抗凋亡活性的评估 表示M11的线条；3)确定M11是否与自身关联 和/或其他Bcl-2家族蛋白以及是否识别的二聚化结构域 可能参与检测到的任何此类关联；4)构建 MHV-68 M11基因缺失重组体(及邻体表达 基因完整)，以确定M11是否是裂解复制所必需的 以及M11是否显著调节细胞的凋亡机制。 对选定的细胞系进行实验性感染。

DESCRIPTION (adapted from the investigator's abstract): The primary objective of this study is to characterize in vitro the regulation of apoptotis by a viral Bcl-2 homolog encoded by the M11 gene of murine gammaherpesvirus 68 (MHV-68). The M11 open reading frame is located within a cluster of viral genes that may be latency-associated in MHV-68. This gene cluster is present in related rhadinoviruses including KSHV and herpesvirus saimiri (HSV), but the presence of a Bcl-2 homolog within this cluster is unique to MHV-68. However, KSHV, HVS, and other gammaherpesvirus genomes including that of EBV also encode Bcl-2 homologous proteins with which M11 is potentially comparable. In the proposed in vitro studies both recombinant virus constructs and M11 expressing cell lines will be used to explore M11 function. Precisely designed MHV-68 recombinants along with conventional expression studies using wild type virus are also expected to provide initial insight into the expression of M11 in specific cell types in vitro providing preliminary data that may be pursued if warranted in future in vivo studies beyond the scope of the current proposal. Specific goals of the proposed study include: 1) Analysis of expression of M11 and adjacent genes during lytic infection of NIH/3T3 cells and during experimental infection of selected murine B-lymphocyte and macrophage cell lines; 2) Assessment of putative M11 anti-apoptotic activity in selected cell lines expressing M11; 3) Determination of whether M11 associates with itself and/or other Bcl-2 family proteins and whether recognized dimerization domains may be involved in any such associations that are detected; 4) Construction of MHV-68 recombinants with the M11 gene deleted (and expression of neighboring genes intact) to determine whether M11 is essential for lytic replication in vitro and whether M11 significantly regulates apoptotic mechanisms during experimental infection of selected cell lines.