GENETICS OF PRIMATE 'D' TYPE RETROVIRUSES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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。这个项目的具体目标包括：(1)确定GAG与细胞骨架马达机制的相互作用，(2)确定病毒和细胞组件在衣壳从组装部位到质膜的运输中的作用，(3)定义衣壳与细胞的ESCRT机制的相互作用，以及(4)研究与病毒萌发有关的多花柱转换机制。今年，我们开发了表达前病毒的功能性Gag-GFP，以及允许我们跟踪病毒衣壳组装和运输的Env(糖蛋白)-Cherry结构。并通过视频显微镜实时观察衣壳与包膜糖蛋白之间的相互作用。我们已经证明，衣壳在细胞内的转运动力学依赖于细胞中病毒糖蛋白和细胞骨架元件的存在。微管的破坏导致衣壳运输的快速停止，并通过一种未知的机制缓慢恢复。 我们先前已经证明M-PMV可以被新世界猴TRIM-5a蛋白所抑制。我们现在已经在基因组序列水平上表征了TRIM5/6/22/34基因簇中基因的差异进化，(2)这些基因的差异调控是这些变化的结果。 我们发现，与TRIM6和TRIM34相比，TRIM5和TRIM22基因的基因组序列有更多的变化。这种变化的主要特征是将转座元件固定在这些基因的内含子区域。TRIM22中的一个这样的LTR在东半球灵长类动物(人类和猕猴)中提供了TRIM22对P53的反应性，而缺乏TRIM22的新大陆灵长类动物(松鼠猴子)无法在P53刺激后上调TRIM22。因此，TRIM5和TRIM22内含子中固定的LTrs可能允许这些基因的不同表达模式。

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.