GENETICS OF PRIMATE 'D' TYPE RETROVIRUSES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 了解逆转录病毒的功能对于寻求治疗或预防艾滋病毒至关重要。 该项目的具体目标包括：（1）确定Gag与细胞骨架运动机制的相互作用，（2）确定病毒和细胞组分在衣壳从组装位点转运到质膜中的作用，（3）定义衣壳与细胞的ESCRT机制的相互作用，以及（4）今年，我们已经开发了一种功能性的Gag-GFP表达前病毒，以及一种Env表达前病毒，（糖蛋白）-Cherry构建体，使我们能够通过视频显微镜实时跟踪病毒衣壳组装和运输，以及衣壳和包膜糖蛋白之间的相互作用。我们已经表明，衣壳的细胞内转运动力学依赖于细胞中病毒糖蛋白和细胞骨架元素的存在。 微管的破坏导致衣壳运输的快速停滞，通过未知的机制缓慢恢复。 我们以前表明，M-PMV可以被新世界猴TRIM-5a蛋白抑制。我们现在已经在基因组序列水平上表征了TRIM 5/6/22/34基因簇中基因的差异进化，（2）证明了这些基因的差异调节是这些变化的结果。 我们已经发现，与TRIM 6和TRIM 34相比，TRIM 5和TRIM 22基因的基因组序列存在显著更多的变化。这种变化的主要特征是转座因子固定到这些基因的内含子区域。TRIM 22中的一个这样的LTR在旧世界灵长类动物（人类和恒河猴）中提供TRIM 22的p53响应性，而TRIM 22缺乏该LTR的新世界灵长类动物（松鼠猴）在p53刺激后不能上调TRIM 22。因此，已经固定在TRIM 5和TRIM 22的内含子中的LTR可以允许这些基因的差异表达模式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Understanding the function of retroviruses is critical in seeking treatment or prevention of HIV. The specific aims for this project include: (1) defining the interaction of Gag with the cytoskeletal motor machinery, (2) determining the role of viral and cellular components in the transport of capsids from the site of assembly to the plasma membrane, (3) defining the interactions of capsids with the ESCRT machinery of the cell and (4) investigating the myristyl switch mechanism that is involved in virus budding.This year we have developed a functional Gag-GFP expressing provirus, as well as an Env (glycoprotein)-Cherry construct that allows us to follow virus capsid assembly and transport, and interactions between capsids and the envelope glycoprotein in real-time by video microscopy. We have shown that the kinetics of intracellular transport of capsids is dependent on both the presence of the viral glycoprotein and cytoskeletal elements in the cell. Disruption of microtubules results in a rapid arrest in capsid transport, that slowly recovers through an unknown mechanism. We showed previously that M-PMV can be inhibited by new world monkey TRIM-5a proteins. We have now characterized the differential evolution of the genes in the TRIM5/6/22/34 gene cluster at the genomic sequence level, (2) demonstrated differential regulation of these genes as a consequence of the changes. We have found that, there is significantly more change in the genomic sequences of TRIM5 and TRIM22 genes compared to TRIM6 and TRIM34. The main characteristic of this change is fixation of transposable elements into the intronic regions of these genes. One such LTR in TRIM22 provides for p53 responsiveness of TRIM22 in Old World primates (humans and rhesus macaques), while New World primates (squirrel monkeys) whose TRIM22 lack this LTR are unable to upregulate TRIM22 following p53 stimulation. Thus the LTRs that have become fixed in the introns of TRIM5 and TRIM22 may allow for differential expression patterns of these genes.